∗

Malgorzata Kesik-Brodacka
Progress in biopharmaceutical development

Department of Bioengineering, Institute of Biotechnology and Antibiotics,

Warsaw, Poland

Abstract
Since its introduction in 1982, biopharmaceutical drugs have group of biopharmaceuticals are biosimilars. Their
revolutionized the treatment of a broad spectrum of diseases introduction in the European Union and, recently, the Unites
and are increasingly used in nearly all branches of medicine. States markets will reduce the costs of biopharmaceutical
In recent years, the biopharmaceuticals market has developed treatment. This review highlights recent progress in the field
much faster than the market for all drugs and is believed to of biopharmaceutical development and issues concerning the
have great potential for further dynamic growth because of the registration of innovative biopharmaceuticals and biosimilars.
tremendous demand for these drugs. Biobetters, which The leading class of biopharmaceuticals, the current
contain altered active pharmaceutical ingredients with biopharmaceuticals market, and forecasts are also discussed.
enhanced efficacy, will play an important role in the 
C 2017 International Union of Biochemistry and Molecular Biology, Inc.

development of biopharmaceuticals. Another significant Volume 65, Number 3, Pages 306–322, 2018

Keywords: biobetter, biopharmaceutical, biosimilar, drug market,

monoclonal antibodies, recombinant vaccines

1. Biopharmaceuticals Biopharmaceuticals have many advantages. For instance,

they target only specific molecules, rarely causing the side
Biopharmaceuticals represent some of the best accomplish-
effects associated with conventional small-molecule drugs [2].
ments of modern science. These drugs are increasingly being
Additionally, compared with conventional drugs, biopharma-
used in practically all branches of medicine and have become
ceuticals exhibit high specificity and activity [3]. The application
one of the most effective clinical treatment modalities for a
of biopharmaceuticals has facilitated the treatment of patients
broad range of diseases, including cancers and metabolic
who respond poorly to traditional synthetic drugs.
disorders.
The term “biopharmaceuticals” was coined in the 1980s
1.1. Biopharmaceuticals and synthetic drugs
and refers to pharmaceuticals produced in biotechnological
Biopharmaceuticals differ from synthetic drugs in all respects.
processes using molecular biology methods. Thus, this group of
The differences between these two categories of drugs include
products was distinguished from the broad category of biolog-
the nature of the product, the source of the active agent, bioe-
ics, which are pharmaceuticals produced using conventional
quivalence criteria, identity, structure, manufacturing methods,
biological methods [1].
composition, dosing, formulation, handling, intellectual prop-
erty rights, legal regulations, and marketing [1].
Biopharmaceuticals are produced in living cells, whereas
synthetic drugs are the products of chemical processes.
Abbreviations: ADC, antibody–drug conjugate; bsAbs, bispecific antibody;
CFPS, cell-free protein synthesis; CRISPR, clustered regularly interspaced Most synthetic drugs are small molecules. For example,
short palindromic repeats; EPO, erythropoietins; EMA, European Medicines a molecule of acetylsalicylic acid is composed of 21 atoms.
Agency; FDA, U.S. Food and Drug Administration; G-CSF, granulocyte In contrast, biopharmaceuticals are typically 100–1000 times
colony stimulating factor; mAb, monoclonal antibody; scFv, single-chain larger [4]. The active pharmaceutical ingredient of such a drug
variable fragment; TNF, tumor necrosis factor; VLPs, virus like particles..
∗ Address
may contain 2000–25,000 atoms. Biopharmaceuticals are also
for correspondence: Malgorzata Kesik-Brodacka, PhD,
structurally much more complex because of the formation of
Department of Bioengineering, Institute of Biotechnology and Antibiotics,
Staroscinska 5, 02-516 Warsaw, Poland. Tel: +48 22 378 62 28; Fax: +48 polymeric chains, which vary greatly in their structure.
22 848 33 32; e-mail: [email protected] The purity of the active ingredient in a pharmaceutical
Received 1 June 2017; accepted 26 September 2017 drug and the composition of the final product can be verified
DOI: 10.1002/bab.1617 relatively easily. Highly pure chemical substances from various
Published online 2 November 2017 in Wiley Online Library sources, including those composed of a mixture of isomers, can
(wileyonlinelibrary.com) be generally considered similar or even identical for practical

306
purposes [1]. The situation is different for biopharmaceuticals. biopharmaceutical can be duplicated completely, even if the
Because of the biological differences between the expression expression systems used in its manufacture are identical
systems and the conditions of the applied manufacturing (e.g., mammalian cells or bacteria). Biosimilars may potentially
process, a certain degree of variability may occur, even between differ from the innovative reference drugs in their glycosylation
different batches of the same product [5]. Therefore, batch- pattern or the electrical potential of the active pharmaceu-
to-batch variations must be monitored to ensure conformance tical ingredient. These differences may influence the quality,
within a specific range. The properties of active pharmaceutical strength, and safety of the drug [21, 22]. As a result, phar-
ingredients in biopharmaceuticals, other than their primary macokinetic and pharmacodynamic properties of biosimilars
structure (e.g., the amino acid sequence), significantly depend and reference biopharmaceuticals may also differ. However,
on the manufacturing method. For this reason, it is assumed the tremendous progress made in bioproduction and analyt-
that “the process defines the product” for biopharmaceuticals ical methods has made it possible to produce proteins and
[6, 7]. glycoproteins that are similar to the reference product [23].
Other characteristics of biopharmaceuticals that dis-
tinguish them from synthetic drugs are their sensitivity to
degradation in the alimentary system and limited penetrability 1.2.1. Registration of biosimilars and generics
through the intestinal epithelium [8]. As a result, they are The fact that biosimilars are not generics affects their registra-
typically administered parenterally via direct injection rather tion procedures. The registration requirements for biosimilars,
than orally [3]. Biopharmaceuticals also require complex although less stringent than those for innovative biopharma-
stabilization systems because of their temperature sensitivity. ceuticals, are much stricter than those imposed on generics.
Unlike synthetic drugs, biopharmaceuticals exhibit much Biosimilars are registered based on their confirmed biosimilar-
more complex mechanisms of action. For example, interferon ity to the corresponding, previously registered innovative drug.
affects the expression of more than 40 genes. Such exten- These pharmaceuticals may be registered after the presenta-
sive complexity often makes it difficult to determine these tion of the required documentation, including a comparison
pharmaceuticals’ complete mechanisms of action [9]. with the reference drug, or after the presentation of complete
Moreover, in contrast to synthetic drugs, biopharmaceuti- documentation, such as that required for an innovative drug.
cals are potentially immunogenic. Even relatively small differ- The relevant European guidelines [17] were adopted in
ences in the structure of the active ingredient may considerably 2005, and several years later, in 2010, a similar set of guidelines
affect the immunogenicity of a drug [10–12]. Process-related was introduced in the United States [24].
impurities may also be immunogenic [13, 14]. The first biosimilar, somatotropin (brand name Omni-
trope), was registered in the EU in 2006. Currently, in the EU,
1.2. Generics versus biosimilars 23 biosimilars have been registered with the EMA, includ-
Generics are defined as drugs that are equivalents of the ing five erythropoietins (EPO) used to treat anemia caused
innovative reference drugs containing the same active pharma- by dialysis and chemotherapy, seven filgrastim-granulocyte
ceutical ingredient. The term refers to substitutes for synthetic colony stimulating factors (G-CSF) administered to treat leu-
drugs. For these drugs, because of their characteristics, the copenia caused by chemotherapy, one human growth hormone
production of a preparation containing an exact copy of the administered to treat growth disorders, two folliculotropic
active pharmaceutical ingredient is relatively rapid, simple, hormones used to treat fertility disorders, two insulin glargine,
and inexpensive. According to the data of the American Federal two enoxaparin sodiuman−anticoagulant used to prevent blood
Trade Commission, the development of a generic drug requires clots, and four antibodies, including infliximab and etanercept
3–5 years and costs $1–5 million [15]. Additionally, a generic [25]. The next wave of biosimilars is expected to be monoclonal
version may be 80–90% cheaper than the innovative reference antibodies [26]. The first biosimilar monoclonal antibody (in-
drug [16]. fliximab) was registered in the EU in 2013. Infliximab is an
The term “generic” is not used in reference to biophar- antibody against tumor necrosis factor (anti-TNF) and is used
maceuticals. The European Medicines Agency (EMA) decided to treat autoimmune disorders, such as rheumatoid arthritis
that the term “biosimilars” should be used in the European and Crohn’s disease. This drug was registered as two individ-
Union (EU) to refer to biological medical products containing ual products under the brand names Inflectra and Remsima
a version of the active pharmaceutical ingredient found in because the active pharmacological agent produced by one
previously registered reference biological medicinal products company is converted into the final drug by two independent
[17, 18]. The U.S. Food and Drug Administration (FDA) use manufacturers.
the term follow-on biologics. Moreover, both of these agencies In the United States, the first biosimilar, filgrastim-sndz
determined that biosimilars may have different actions than (brand name Zarxio), was approved by FDA in March 2015
the reference drug [19, 20]. [27]. Zarxio is produced by the European company Sandoz and
These differences are related to the application of dif- is a biosimilar of filgrastim (brand name Neupogen), which
ferent expression systems and different manufacturing and was originally developed and manufactured by Amgen Inc.
purification processes in the production of biosimilars. No Zarxio is registered for the treatment of the same conditions as

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Neupogen [27]; however, it was approved as a biosimilar, not a the risk of failure during trials is much lower for a biosimilar.
substitute. For this reason, biosimilar drugs may potentially replace the
Considering that the patent protection period has already reference drug on the market after the expiration of its patent
expired or will soon expire for several important biophar- protection. Manufacturers of reference drugs may attempt to
maceuticals (Fig. 1), it might be possible to register other prevent this potential replacement by introducing a biobetter,
biosimilars in the near future. which is an improved version of the reference drug. If the
additional advantages of a biobetter are so significant that
1.3. Biobetters it may be preferable in therapy, then the development of a
Another group of biopharmaceuticals are biobetters. Biobetters biosimilar may not be viable or the introduction of such a drug
are biopharmaceuticals that have been structurally and/or will bring much lower profits. Thus, biobetters may limit the
functionally altered to achieve an improved or different clinical market share for biosimilars. In contrast, the development of
performance, compared to approved reference products [18]. biobetters requires more extensive research than biosimilars,
Biobetters are treated by regulators as different from the greatly increasing the costs of drug development. For this
existing products and, therefore, are evaluated as new drugs, reason, it is crucial for a drug’s success for that drug to have
in a standard approval procedure. therapeutic benefits that are significant enough to justify its
Biobetters represent the next stage in the development broad application, despite its potentially higher price.
of biopharmaceuticals in which proteins were purposefully One example of the competition described above is the
altered equivalents of existing drugs. The changes introduced case of the drugs used to treat chronic myeloid leukemia.
were aimed to improve the proteins, i.e., obtain stronger The reference biopharmaceutical rituximab (brand name
clinical effects, require less frequent administration, achieve MabThera) was patented in 1993 in Europe and 4 years later
better targeting, and/or be better tolerated compared with their in the United States. However, in 2014, 2 years before the
equivalents [28]. For this purpose, these proteins were opti- expiration of rituximab’s patent protection, the companies
mized for favorable biodistribution and pharmacokinetic and producing this drug introduced the biobetter obinutuzumab
pharmacodynamic properties. The introduced modifications (brand name Gazyva/Gazyvarol) into the market. Rituximab is
included changes in the amino acid sequence and the glyco- a monoclonal anti-CD20 antibody and shows greater efficacy
sylation pattern of a given protein. In the case of monoclonal than the reference biopharmaceutical. Additionally, it must be
antibodies, pegylation or combination with a cytotoxic drug are stressed that Gazyva had been registered before the studies on
strategies to enhance their efficacy tor change their half-life. a biosimilar were completed [32].
The first such biopharmaceutical produced with an altered
amino acid sequence was the fast-acting insulin analog Lispro 1.4. Role of cytochrome P450 enzymes in drug
(brand name Humalog), which was registered in the United development
States in 1996. Another example of such an improvement is the Metabolism of drugs is a complex process in which many
extension of the pharmacokinetic half-life of equivalents of rit- different enzymes are involved. Among these enzymes are
uximab, trastuzumab, and bevacizumab, which was achieved cytochromes P450, a large superfamily of ubiquitous heme-
by introducing two or three amino acid mutations in the Fc containing monoxygenases [33, 34]. These enzymes are es-
domain [29]. In this case, the extension of intervals between sential for metabolism of 80% of clinically used drugs. In the
the drug administration or reduction in the dose is expected to drug metabolism, multiple products may be obtained from
lower the treatment costs. the same drug and one drug may be metabolized by more
Another example of a biobetter is the ado-trastuzumab than one cytochrome P450 enzyme. Moreover, each enzyme
emtansine (brand name Kadcyla), an antibody–drug conju- acts on more than one drug. The substances resulting from
gate produced by Roche and an improved equivalent of the a drug metabolism can be biologically active and may cause
trastuzumab antibody (brand name Herceptin) manufactured adverse drug reactions [33]. Therefore, in the drug discovery
by the same company [30]. Kadcyla slows down disease pro- and development process it is of key importance to investigate
gression by almost twofold in patients with HER2-positive the metabolism of drug candidates. This has led to increased
advanced breast cancer, extending the median total survival demand for drug metabolites to facilitate evaluation of their
time by 5.8 months relative to other treatment methods [31]. possible adverse effects in animals and humans as well as
Both biosimilars and biobetters are natural alternatives drug’s efficacy and pharmacokinetics.
for the reference biopharmaceuticals and therefore compete The use of human cytochromes P450 to produce drugs,
for the same market. This competition is influenced by the drug metabolites, and intermediates is mainly limited by their
costs of developing and launching the drug in the market, legal poor solubility, stability, and low coupling [35]. By contrast,
regulations connected with the registration of individual groups P450 BM3, bacterial P450 enzyme from Bacillus megaterium,
of drugs, and the time required to proceed from introduction has been shown to be able to produce drug metabolites typ-
to turnover. As a rule, the development of a biosimilar is ical of the human enzymes with a high coupling efficiency
faster and cheaper than that of a reference drug because a [33, 36]. Owing to P450 BM3 characteristics, multiple protein
biosimilar is an equivalent of an existing drug. Additionally, engineering studies have been performed on this enzyme to

308 Progress in Biopharmaceutical Development

 Dates of patent expiry for the 10 best-selling systems based on fungi (Saccharomyces cerevisiae and Pichia
FIG. 1 biopharmaceuticals in 2016. * Data not available. pastoris), mammalian cells, or insect cell lines. The use of
cell-free expression systems (in vitro systems), which greatly
facilitates modifying synthesis conditions, has also been studied
widen its catalytic abilities [37–40]. Moreover, several con- [47].
structions have been already reported including different The production of biopharmaceuticals in each of the
fusion human P450 enzymes, engineered by connecting the aforementioned systems has advantages and drawbacks.
P450 BM3 reductase domain with human cytochromes P450 For these reasons, many different expression systems are used
3A4, 2C9, 2C19 [41], 2A6, CYP2C6, and CYP4F11 [42], monkey based on the specific properties of a given recombinant protein.
2C20 [43], and dog CYP2D15 [44, 45]. Also the catalytic perfor-
mance of one of the created chimeric proteins was improved in 2.1. Mammalian expression system
terms of coupling efficiency and enzyme turnover by engineer- Mammalian expression systems are generally the preferred
ing the loop connecting the two domains [46]. The published platform for manufacturing biopharmaceuticals. In recent
results were an important factor in engineering catalytically years, a steady increase in the use of these expression systems
self-sufficient human P450 for applications in biocatalysis [46]. has been observed. This is because of the growing interest
in the production of large, complex molecules that require
specific posttranslational modifications (most notably glycosy-
lation) that occur only in mammalian expression systems [48].
2. Systems for the Production of
Additionally, in the case of mammalian cell lines and animal
Biopharmaceuticals cell lines in general, most recombinant proteins can be secreted
Unlike synthetic drugs, the active pharmaceutical ingredients in and do not require cell lysis to extract with subsequent protein
biopharmaceuticals include recombinant proteins and nucleic refolding (as is the case with bacteria) [48].
acids. Currently, the vast majority of commercially available However, protein production in cell lines raises potential
biopharmaceuticals contain recombinant proteins as their safety concerns due to the possibility of contaminants with
active pharmaceutical ingredient. These proteins are produced animal viruses. Other drawbacks of protein production in
in prokaryotic systems, mainly Escherichia coli, or eukaryotic cell lines include the complex nutritional requirements, slow

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growth and fragility, and a relatively high production time and posttranslational modifications that occur within the cells often
cost [49]. Currently available mammalian expression systems lead to the production of undesired hypermannosylation [56],
include Chinese hamster ovary cells, rodent cell lines (e.g., which can result in altered protein binding activity, and po-
NS0, BHK, and Sp2/0), and human cell lines (e.g., HEK293, tentially yield an altered immunogenic response in therapeutic
PER.C6, HT-1080, and CAP) [50]. Among available mammalian applications. In P. pastoris, oligosaccharides have much shorter
cell lines, Chinese hamster ovary cell line is the primary choice chain lengths and the strain has been reported to produce com-
for recombinant protein production, with 7 of the top 10 best- plex, terminally sialylated or “humanized” glycoproteins [58].
selling biopharmaceuticals from 2016 being produced in these P. pastoris is an expression system that is appreciated for its
cells. In general, the number of recombinant protein products growth to very high cell densities, for its available strong and
produced in mammalian systems that are approved for use as tightly regulated promoters, and for the possibility to produce
drugs in humans increased over 2010–2014 to approximately gram amounts of recombinant proteins per litre of culture both
60% [49]. intracellularly and in a secretory fashion [59]. However, protein
yields can be remarkably lower, particularly if the expressed
2.2. Bacterial expression system complex proteins are heterooligomers, membrane-attached or
Nevertheless, bacteria remain the dominant expression system, prone to proteolytic degradation [59].
facilitating the production of large quantities of active phar-
maceutical ingredients used in biopharmaceuticals. According
2.4. Insect cell line expression system
to the data provided by BioProcess Technology Consultants, in
Insect cell-based recombinant protein production system rep-
2010, the total production of pure proteins as active pharma-
resents a compromise between bacterial and mammalian
ceutical ingredients in biopharmaceuticals amounted to 26.4
expression systems. Its advantage over the bacterial system
tons. Of this, 68% were produced in bacterial systems and 32%
is that it allows for posttranslational modifications but unlike
in mammalian systems. The predominant group of proteins
mammalian system it does not preserve the original glycosy-
produced in bacteria comprised insulins, and the vast majority
lation pattern. [60]. Another advantage of use of insect cells is
of those produced in mammalian systems were monoclonal
that they are less demanding and they grow to higher densities
antibodies [51].
compared to mammalian cells.
The bacteria of choice for heterologous protein expres-
An insect cell line expression system was used to produce
sion are E. coli. Its attractiveness for industrial applications
Cervarix, a vaccine against certain types of cancer-causing
results from its well-understood genetics, cell biology, and easy
human papillomavirus. This vaccine was approved by the EMA
handling. Expression systems based on E. coli allow for rapid
in 2007.
growth, high product yield, cost-effectiveness, easy process
scale-up, and short turnaround time [52, 53]. The limitations of
this expression host for the production of complex recombinant 2.5. Transgenic animals
biopharmaceuticals include the absence of mammalian-like In addition to commonly used prokaryotic and eukaryotic
posttranslational modifications, such as glycosylation, phos- expression systems, there is an increasing interest in the use of
phorylation, and proteolytic processing [54]. Hence, E. coli is transgenic animals for recombinant protein production. This
the expression host of choice in the biotechnology industry for is due to the low cost of producing large amounts of complex
the large-scale production of small recombinant proteins that proteins in these systems [61, 62]. Transgenic animals offer the
do not require posttranslational modifications [49]. Another opportunity to produce human recombinant proteins with post-
limitation results from E. coli’s inability to produce correct translational modifications that closely match human proteins.
disulfide bonds, protein solubility issues, and the presence of However, there are issues with the generation of transgenic
endotoxins (lipopolysaccharide) [55]. Currently, several strate- founders. Although many strategies have evolved over the past
gies are applied to improve protein expression, such as the use decades, transgenesis in animals is relatively inefficient and
of mutated E. coli strains to promote protein disulfide bond time consuming. Attempts to improve transgenesis by various
formation [52]. methods have had limited success, mainly due to random trans-
gene integration and the control over transgene copy number
2.3. Yeast expression system [63].
Additional favorable microbial recombinant protein production The first biopharmaceutical, ATryn, whose active pharma-
systems are the eukaryotic microorganisms S. cerevisiae and ceutical ingredient was produced in transgenic animals (goats),
P. pastoris [56]. Both of these hosts are capable of producing was launched on the market in 2006 in the European Union
recombinant proteins with proper folding and posttranslational (2009 in the United States). This drug is an anticoagulant and
modifications [57]. Therefore, they are considered better than contains a plasma protein, human alfa antithrombin [64]. Since
prokaryotes where posttranslational modification of the target then, other proteins produced in transgenic rabbits have been
protein is needed. The S. cerevisiae yeast expression system is approved for use. Thus, conestat alfa (brand name Ruconest), a
frequently used due to their rapid growth in protein-free media recombinant analogue of the human esterase inhibitor C1, was
and ability to secrete the product extracellularly. However, approved for the treatment of hereditary angioedema [65]. The

310 Progress in Biopharmaceutical Development

recombinant proteins produced by these animals are secreted costs, small reaction scales, and limited ability to correctly fold
into their milk [66]. proteins containing multiple disulfide bonds.
Currently, because of the significant progress made to
2.6. Plant expression system automate and optimize reaction conditions, cell-free systems
The production of biopharmaceuticals derived from plants have become an attractive protein production platform offer-
has attracted great interest. Transgenic plants have the po- ing several advantages over traditional cell-based expression
tential to become cost-effective systems for the large-scale methods [74]. First, CFPS environment is not limited by the
production of human therapeutic proteins. The use of plants presence of a cell wall or homeostasis conditions to maintain
eliminates potential contamination of the therapeutic drug with cell viability. CFPS enables direct access, and therefore control
animal pathogens, as plant cell cultures are not susceptible to of the translation environment, and manipulation of the re-
mammalian viral pathogens and, conversely, plant viruses do action composition and conditions, which is advantageous for
not infect human cells [67]. Another advantage is that orally the optimization of protein production. As a result, new com-
immunogenic recombinant proteins expressed in an edible ponents can be added/synthesized and maintained at precise
plant may be orally administered without processing, including concentrations [75]. Other advantages of CFPS over cell-based
expensive purification steps [68]. Moreover, plant expression systems include the ability to produce difficult to express pro-
systems are able to produce proteins with complex glycosy- teins, e.g., membrane proteins as well as toxic proteins [76].
lation patterns; however, the glycan structures produced are Unlike systems based on living organisms, it is believed that
significantly different from those produced in humans. the protein synthesis conditions in CFPS are analogous to those
Drawbacks of plant-based expression system for recombi- of chemical reactions, which is promising from the perspective
nant protein production are related to long production timelines of technical scalability.
that render this technology unsuitable for the rapid production Compared to expression methods based on bacterial or
of pharmaceuticals to combat emerging diseases [69]. Another tissue culture cells, CFPS is considerably faster because it does
issue is that current methods in plant biotechnology cannot not require gene transfection, cell culture, or extensive protein
precisely control transgene expression levels in plants in a purification. Moreover, the speed and ease of protein production
consistent manner [70]. in CFPS results from the possibility of direct expression from
As an alternative to using whole plants as bioreactors, PCR-generated templates without requiring fragment cloning.
there has been considerable progress in the use of plant cell Despite of the made progress CFPS still suffers from the lower
cultures, such as carrot suspension culture and tobacco BY-2 yield of target proteins and relatively high cost compared to the
cells [71]. Until now, the main classes of biopharmaceutical other expression systems.
proteins successfully produced and correctly folded in plants Recently, Sutro Biopharma has developed STRO-001, an
have been subunit vaccines and virus-like particles (VLPs), antibody–drug conjugate (ADC). It was developed via propri-
antibodies, and therapeutic enzymes, including several prod- etary cell-free protein synthesis and site-specific conjugation
ucts that have completed phase II trials and are close to platforms, which facilitate multiple rounds of antibody and
commercialization [70]. In 2012, a protein produced in plant ADC optimization. STRO-001 is a novel CD74-targeting ADC
cell lines (carrot root cells) was allowed to enter the drug composed of a p-azido-methyl-phenylalanine-containing anti-
market. This protein, recombinant human glucocerebrosidase CD74 aglycosylated human IgG1 antibody (SP7219) conjugated
(brand name Elelyso), is an active pharmaceutical ingredient to a noncleavable dibenzocyclooctyne-maytansinoid linker-
in the drug used to treat Gaucher’s disease and became the warhead. [77]. STRO-001 has been shown to eradicate tumors
first plant-produced biopharmaceutical approved for human in human xenograft models of non-Hodgkin lymphoma and
administration by the FDA [72, 73]. Since Gaucher’s disease is multiple myeloma diseases. The company plans to submit
a rare disease, treatment of this disease with an orphan drug is an Investigational New Drug application to the FDA at the
costly (US$200,000 annually per patient for life). The use of a end of 2017 and initiate STRO-001 clinical testing in the
carrot cell production system reduced the cost to US$150,000 first quarter of 2018. If it passes the tests, it will be the first
per patient per year [70]. commercial biopharmaceutical produced in a cell-free expres-
sion system, which would prove commercial viability for this
2.7. Cell-free protein synthesis technique.
Cell-free protein synthesis (CFPS), also called in vitro
expression, is an innovative and promising alternative to
the expression of recombinant proteins in living cells. CFPS 3. First Gene Therapies
is the production of recombinant proteins using translation In addition to recombinant proteins, nucleic acids may also
machinery extracted from cells. In this system, the enzymes be biopharmaceuticals’ active pharmaceutical ingredients.
required for the transcription and translation processes are Most studies on gene therapy have focused on the induction
present in a cell extract instead of a live organism. Several or inhibition of cellular processes underlying diseases. Gene
obstacles initially limited CFPS as a protein production tech- therapy is based on the introduction of genetic material into an
nology, including low protein production rates, high reagent organism or patient, either directly or using viruses.

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A breakthrough in the biopharmaceutical sector was the approval is expected by the end of 2017. If the FDA accepts the
registration of a DNA-based drug (the first gene therapy). The recommendation, this treatment will be the first gene therapy
first drug used in gene therapy (Alipogene tiparvovec, brand to ever reach the market in the United States.
name Glybera) was approved for use in the EU in 2012 [78]. This While most gene therapy trials have addressed cancer,
therapy compensates for the lipoprotein lipase deficiency found a significant number of gene therapy trials have targeted
in rare hereditary disorders that leads to severe pancreatitis. rare inherited monogenic diseases (10.5% of all gene therapy
Unfortunately, Glybera administration provides only temporary trials) [86]. Monogenic diseases receive much attention as they
relief [51]. Initially, the cost of a single treatment was estimated hinge on the conceptually simple idea that diseases caused
at $1.6 million [79]. In 2015, this figure decreased to $1 million by a known single gene defect could be potentially cured by
[80], yet this therapy remains the most expensive in the world. the insertion and expression of a single correct copy of the
Glybera failed to achieve recognition of benefit in two EU mutant or deleted gene in host cells. One monogenic disease
countries (France and Germany) and is unlikely to ever be that may be curable in the near future using gene therapy is
commercialized in other European countries. Moreover, plans the inherited bleeding disorder hemophilia B. After receiving
for the commercialization of this therapy in the United States a single dose of an experimental gene therapy (SPK-9001) in a
have been abandoned. Thus, over 4 years after the European clinical trial, patients with hemophilia produced near-normal
regulatory approval, the first commercial gene therapy drug levels of clotting factor IX, allowing them to stop clotting factor
has been used in only one case [81]. infusions and to pursue normal daily life activities without
A few years earlier, in 2003, another drug used in human disabling bleeding episodes [88].
gene therapy, Gendicine, was approved in China for the treat- Over 77% of all gene therapy clinical trials performed to
ment of head and neck squamous cell carcinoma [82]. In 2011, date are phase I or I/II. Ninety-three gene therapy clinical trials
Neovasculgen was registered in Russia as a first-in-class gene- were in phase III. It has been predicted that by 2020, some 5–10
therapy drug for the treatment of peripheral artery disease, gene therapies will be available. The first gene therapies are
including critical limb ischemia. expected to be used to treat a rare form of blindness, Leber’s
Recently, three other gene therapies (Imlygic, Strimvelis, congenital amaurosis, sickle-cell anemia, beta-thalassemia,
and Invossa) were approved. Imlygic (Talimogene laher- and a spectrum of rare cancers and genetic diseases [89, 90].
parepvec) was approved by the EMA and the FDA in October However, given the lessons learned from the development of
2015. It is a modified form of the herpes simplex virus type 1 for Glybera gene therapy, the high costs and the temporary nature
the local treatment of unresectable cutaneous, subcutaneous, of its beneficial effects, we will likely not see the widespread use
and nodal lesions in patients with recurrent melanoma after of such therapies in the near future. Nevertheless, the changes
initial surgery [83]. brought about by the introduction of gene therapies will spark
In April 2016, the EMA approved the first ex vivo stem cell an era of targeted and personalized treatment [90].
gene therapy (Strimvelis) with indications to treat patients with Recent research shows that the development of gene
adenosine deaminase-deficient severe combined immunodefi- therapy can be accelerated by a new, revolutionary genome-
ciency [84]. Strimvelis consists of autologous gene-corrected editing tool: clustered regularly interspaced short palindromic
hematopoietic stem cells and is prepared from the patient’s own repeats (CRISPR) [91]. It was successfully used for in vitro
bone marrow hematopoietic stem cells, which are genetically CRISPR-based genome editing to correct defective genotypes
modified using a gamma-retroviral vector to insert a functional [91]. Moreover, several studies have also shown that CRISPR
copy of the adenosine deaminase gene [85]. therapies can be successfully implemented in vivo [92]. There
In July 2017, South Korea’s Ministry of Food and Drug are currently two clinical trials involving CRISPR-Cas9 for
Safety approved the country’s first gene therapy drug. The targeted cancer therapies that have been approved in China
drug, Invossa, is the world’s first cell-medicated gene therapy and the United States [91]. On October 2016 as a part of
for osteoarthritis. Invossa uses allogeneic human cartilage cells clinical trials, a group of Chinese researchers used immune
engineered to express transforming growth factor TGF-ß1. The cells that contain a gene (PD-1) edited using this technique.
drug was approved for sales in the domestic market. The cells were removed from the blood of a person with lung
cancer, the gene was disabled, and the cells were injected
3.1. The future of gene therapy back into the patient. In 2017, another Chinese group plans to
Between 1989 and April 2017, 2,463 gene therapy clinical start three clinical trials of drugs developed using the CRISPR
trials have been completed, are ongoing or have been approved technique. These therapies target bladder, prostate, and renal
worldwide [86]. So far, most of them have been aimed at the cell cancers [93]. In June 2016, the U.S. National Institutes of
treatment of cancer (64.4% of all gene therapy trials) [86]. Health approved the first CRISPR clinical trial in the United
In July 2017, the FDA advisory panel recommended States to help augment cancer therapies.
approval of tisagenlecleucel-T, a chimeric antigen receptor As CRISPR-based treatments have made enormous
T-cell therapy. The treatment would be directed for children progress since their beginning only a few years ago,
and young adults with B-cell acute lymphoblastic leukemia there is great hope that this tool will strongly accelerate
[87]. Chimeric antigen receptor T-cell therapy treatment the development of gene therapies. Nonetheless, more

312 Progress in Biopharmaceutical Development

investigations are needed to fully harness the power of this a rat-mouse hybrid IgG2 monoclonal antibody produced in a
technique. rat-mouse hybrid cell line (hybridoma), was the first approved
bsAb. This antibody is used in the intraperitoneal treatment of
malignant ascites. Catumaxomab was registered in 2009 in the
4. Therapeutic Antibodies EU and is in clinical trials in the United States. The other bsAb,
Monoclonal antibodies (mAbs) are the largest class of bio- Blinatumomab (trade name Blincyto), was registered in 2014
pharmaceuticals and are currently utilized in therapies for by FDA for the treatment of relapsed or refractory Ph-negative
cancer, inflammatory diseases, cardiovascular diseases, organ acute lymphoblastic leukemia in adults. Blinatumomab is
transplantations, infections, respiratory diseases, and ophthal- currently undergoing multiple phase II and phase III clinical
mologic diseases. This group of biopharmaceuticals includes trials for other B-cell related malignancies [104]. Currently,
mAbs and derivative antibodies, such as bispecific antibodies there are two bsAbs (Emicizumab and MEDI-565) in phase III
(bsAbs), antibody–drug conjugates, radiolabeled antibody con- clinical trials [103]. Emicizumab is a bsAb for prophylactic use
jugates, antigen-binding fragment Fab, and Fc-fusion proteins to reduce the number of bleeding episodes in patients with
[94]. hemophilia A and factor VIII inhibitors. MEDI-565 (also known
The availability of fully human and humanized mAbs as MT111 and AMG 211) is a bsAb for the treatment of patients
increased the therapeutic efficacy for oncology and hema- with cancers expressing carcinoembryonic antigen.
tooncology and for inflammatory and autoimmune diseases. Currently, there are increased numbers of clinical trials
The first monoclonal antibody–based biopharmaceutical was being performed on novel, bsAb-based drugs. Therefore, the
muromonab-CD3 (sold under the brand name Orthoclone introduction of new drugs of this type on the market is expected
OKT3), which is administered during acute kidney transplant in the near future.
rejection [95]. This drug was registered in 1986; however, the
dynamic development of the antibody market began in the 4.2. Radiolabeled antibody conjugates, antibody–drug
late 1990s, when the first chimeric mAb was registered. As conjugates
of March 2017, in the EU and the United States, a total of 71 Another group of monoclonal antibody-based drugs includes
monoclonal antibody-based drugs have been registered [96]. cancer antigen-specific mAbs conjugated with isotopes emitting
Currently, fully human antibodies are growing as a proportion radiation or a highly potent drug. Both of these strategies
of mAbs in the clinic. In 2002, the FDA approved the first fully facilitate the specific destruction of cancer cells. Currently,
human mAb, adalimumab. Since that time, approximately 40% two radio-immunoconjugates are registered for the treatment
of all of the marketed mAbs are fully human. of non-Hodgkin’s lymphoma, 131I-Tositumab (brand name
Bexxar) and 90Y-ibrytumomab tiuksetanem (brand name
4.1. Bispecific antibodies Zevalin) [105, 106].
Most registered antibodies are monospecific antibodies that are In recent years, ADCs have become powerful tools in the
capable of interacting with a single target. However, complex treatment of cancer. An ADC is a bioconjugate that contains a
diseases, such as cancers or inflammatory disorders, are fre- mAb that specifically binds to a tumor virus surface antigen
quently multifactorial in character. In these cases, the inhibition and a highly potent drug that is attached to the antibody
of many different pathogenic factors and signaling pathways via cleavable or noncleavable linkers. This design ensures
may enhance the therapeutic efficacy. For this purpose, bsAbs specificity and efficacy in targeting cancer cells and allows the
were designed [97]. These antibodies are artificial proteins healthy tissues to remain generally unaffected. Recently, the
composed of fragments of two different monoclonal antibodies FDA approved two such drugs, brentuximab vedotin (brand
and thus bind to two different types of antigens. bsAbs are most name Adcetris) and ado-trastuzumab emtansine (brand name
commonly used in cancer immunotherapy, where they simul- Kadcyla).
taneously bind to two targets, e.g., a tumor cell and cytotoxic In 2015, more than 50 new ADCs were in clinical trials all
cells (via a receptor, such as CD3) [98, 99]. bsAbs efficiently over the world [107]. As of January 2017, there are 37 ADCs
stimulate the host immune system, facilitating the destruction in phase I trials. Three ADCs entered phase I/II development in
of cancer cells [100]. 2016, increasing the total number of ADCs in this stage to eight.
Initially, bsAbs were generated via the chemical conjuga- Four ADCs (AGS-16C3F, Anetumab Ravtansine, SAR566658,
tion of two different antibodies, producing a single molecule and Rova-T) progressed toward phase II, yielding 11 phase II
equipped with four antigen-binding regions (four Fab frag- ADCs. Two drugs (IMGN853 and SGN-CD33A) entered phase III
ments), with each of the fragment pairs binding a different trials, which doubled the number of ADCs in this clinical phase
molecule [101]. Another approach to generating bsAbs is the [108]. In August 2017, FDA approval could become a reality
fusion of two hybrids, producing antibodies with differing for inotuzumab ozogamicin [108], the anti-CD22 antibody–
specificities. Currently, more than 50 different technological drug conjugate for the treatment of patients with relapsed or
platforms are available for bsAbs production [102]. refractory acute lymphoblastic leukemia.
So far, the FDA has approved two bsAbs, catumaxomab and Overall, it is estimated that approximately 10 new ADCs
blinatumomab [103]. Catumaxomab (brand name Removab), will come onto the clinical market in the next decade [107].

Biotechnology and Applied Biochemistry 313

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Despite the tremendous progress in ADC development, further mAbs against the antigens present in specific pathological cell
studies are necessary to enhance their safety and efficacy [109]. types are increasingly being used in both therapy and clinical
A major challenge in this regard is ensuring the homogeneity diagnostics. Because of the large number of mAb candidates
of ADC molecules [110]. undergoing evaluation in late-stage clinical studies (over 50 as
of December 2016), a trend toward first marketing approvals
4.3. Single-chain variable fragment of at least six to nine mAbs per year is expected to be observed
An important focus of studies on mAbs relates to the reduction in the near-term. This prediction indicates that the market for
of the active particle size to enhance antibody penetration mAbs is likely to develop dynamically, which will be manifested
following their administration and facilitate their production. in further increases in the sales of these drugs.
This line of research addresses the development of single-chain
variable fragments (scFvs).
5. Vaccines
4.4. Glycoengineered monoclonal antibodies Another potential area of biopharmaceutical growth is vac-
Another technological innovation was the development of cine development. Any given vaccine may be classified as
the first glycoengineered mAb, afutuzumab (brand name biopharmaceutical if molecular biology methods are used in
Gazyva), which was registered in 2013 in the United States its development. An example could be live attenuated vaccines
[111]. The introduced modifications enhanced the antibody- where recombinant DNA technology was used to alter the
dependent cell-mediated cytotoxicity. This antibody is an pathogen’s genome. Another group of vaccines that can be
immunomodulator used to treat lymphoid malignancies [112]. classified as biopharmaceuticals are subunit vaccines, which
4.5. The future of therapeutic antibodies are based on specific, highly purified recombinant protein
Currently, most registered mAb-based drugs are used to antigens.
treat cancers and autoimmune disorders. According to World 5.1. Subunit vaccines
Health Organization estimates, the number of new cancer Subunit vaccines contain only defined antigens instead of whole
cases will increase to 27 million in 2030 [113] because of the pathogens, and, therefore, their application does not introduce
growing number of elderly individuals. Taking into account the risk of infection. However, a major challenge for current
these epidemiological data and the resulting huge demand subunit vaccine development is the fact that many new subunit
for anticancer therapy, it is expected that anticancer drugs vaccines are poorly immunogenic and mobilize insufficient
will be the leading group among all registered drugs. This immune responses for protective immunity. Therefore, effective
is also confirmed by data on the number of mAbs that are adjuvants are needed to enhance, direct, and maintain the
entering clinical trials. During 2014–2016, pharmaceutical immune response to vaccine antigens [116]. New adjuvants are
companies initiated the first in-human studies for an average of designed to not only boost immunological response but also to
approximately 80 mAbs-based therapeutics annually, of which increase cross-protection against different strains or variants of
more than 60% were designed to treat cancer [114]. In 2016, the same pathogen. The new generation of rationally designed
antibodies for cancer represented approximately 55% of the vaccine adjuvants target specific innate immune receptors such
overall clinical pipeline of therapeutic antibodies. Anticancer as toll-like receptor (TLR) 4 and TLR9. These novel adjuvants
mAbs that entered the clinical pipeline in 2017 include two reached human clinical trials stage [117]. Several studies are
(CX-072 and KN035) that target the programmed death-1 also conducted on the synergistic effects of different adjuvants
receptor ligand, and one each (CBT-501 and FLYSYN) that to identify new beneficial effects of vaccine efficiency [118].
target programmed death-1 receptor and Fms-like tyrosine Another crucial aspect of development of next-generation
kinase, respectively [114]. Among the five mAbs that have vaccines is the optimal presentation of the antigen to the
recently entered late-stage clinical studies, three (utomilumab, immune system to achieve desirable immune response. In the
isatuximab, and SHR-1210) are being evaluated as treatments quest for novel and effective presentation methods as well
for cancer. The other two mAbs, crizanlizumab and olokizumab, as delivery strategies, VLPs offer several promises. VLPs can
are being studied in patients with sickle cell disease (for the elicit strong T and B cell immune responses because they
prevention or reduction of the occurrence of pain crises) and contain repetitive displays of viral surface proteins that present
rheumatoid arthritis, respectively [115]. conformational viral epitopes. VLPs are not infectious but have
Extensive research efforts are currently focused on the similar properties to virions, enabling them to be used as both
development of improved antibodies against known molecular particulate carriers and adjuvants in vaccine development.
targets associated with asthma, leukemia, nonsmall-cell lung VLPs were successfully used in approved vaccines for hepatitis
carcinoma, and multiple sclerosis. In the near future, next- B and human papillomavirus.
generation antibodies with improved properties, including
ADCs and bsAbs, are expected to gain popularity as biobetter 5.2. New technologies in vaccine development
antibody therapeutics [103]. Another important subject in biopharmaceutical development
Additionally, studies are being conducted to develop mAb is the emergence of new technologies, which have the potential
biosimilars that are already used in therapy. High-specificity to revolutionize the vaccine field. These technologies include

314 Progress in Biopharmaceutical Development

reverse vaccinology, structural vaccinology, and synthetic 5.3.1. HIV
vaccines. Two HIV vaccines, HVTN 702 and Ad26, are currently part
of efficacy clinical trials in humans. The HVTN 702 vaccine
5.2.1. Reverse vaccinology regimen consists of two experimental vaccines, a canarypox
The concept of reverse vaccinology involves using bioinformat- vector-based vaccine called ALVAC-HIV and a two-component
ics tools to screen the entire genome of a pathogen to identify 120 HIV glycoprotein subunit vaccine with MF59 adjuvant to
genes encoding proteins with the attributes of good vaccine enhance the body’s immune response to the vaccine. The results
targets. Current reverse vaccinology approaches include com- of HVTN 702 clinical trials are expected in late 2020 [125]. The
parative in silico analyses of multiple genome sequences, second study (Ad26) is based on “mosaic” vaccines designed to
which enable the identification of conserved antigens within induce immunological responses against a wide variety of HIV
a heterogeneous pathogen population and the identification subtypes responsible for HIV infections globally. The vaccine
of antigens present in pathogenic but not commensal strains uses a strain of adenovirus serotype 26 as a vector to deliver
[119]. Moreover, transcriptomic and proteomic data sets are in- three (trivalent) or four (tetravalent) mosaic antigens for HIV
tegrated into a selection process that allow for the accelerated variant genes and Clade C 140 HIV glycoprotein adjuvanted
identification of vaccine targets to be tested in animal models. with aluminum phosphate. The results from these clinical trials
Reverse vaccinology was successfully applied against serogroup are expected in late 2017 [126].
B meningococcus [120]. This technology was also used in ad-
vanced preclinical and clinical vaccine studies against several 5.3.2. Tuberculosis
pathogens, including those resistant to antibiotics [121]. Thirteen different tuberculosis vaccine candidates are currently
in clinical trials; eight of them are subunit vaccines, six of which
5.2.2. Synthetic vaccines contain or express one of Mycobacterium tuberculosis antigen
Synthetic vaccines were developed to accelerate vaccine avail- 85 complex proteins (either the Ag85A or Ag85B) [127]. The
ability for future pandemics. This technology enables the rapid current test evaluates the potency of new candidates in animal
generation of vaccine viruses from sequence data. Dormitzer models by measuring the reduction in the bacillary load in
et al. [122] used an enzymatic assembly from chemically the lungs during the acute phase of the infection. However,
synthesized oligonucleotides and improved in vitro error cor- so far, none of the candidates has been able to prevent the
rection for the rapid, accurate gene synthesis of two major establishment of the infection [128].
influenza virus surface glycoproteins (hemagglutinin and
5.3.3. Malaria
neuraminidase). This synthetic approach allowed for the de-
Substantial progress has been made in the development of
velopment of vaccine seeds in a matter of days instead of the
malaria vaccines during the past decade. In 2015, recombi-
typical 2–3 months needed when conventional technologies are
nant protein-based malaria vaccine, RTS,S/AS01 (trade name
used.
Mosquirix), has received a positive opinion from the EMA [129].
RTS,S/AS01 is the world’s first licensed malaria vaccine.
5.2.3. Structural vaccinology
Structural vaccinology is emerging as a promising platform for 5.3.4. Universal flu vaccine
the identification of effective protective antigens to facilitate The progress in the field of biopharmaceuticals may enable
the development of optimized and possibly broadly protective development of a universal flu vaccine. Such vaccine, in contrast
vaccines. In this technology, the domains within an immuno- to currently available vaccines, would be able to provide long
genic protein that contain epitopes inducing protective immune lasting and broad protection against flu infections [130]. Some
responses are identified and expressed in a recombinant form. very promising results were obtained from the structure-
These domains can be used as potent immunogens devoid of based development of an influenza virus H1 hemagglutinin
the regions of the immunogenic protein that are irrelevant from stem-only immunogen. The immunogen confers heterosubtypic
a vaccine standpoint [123]. Recently, it was shown that, with protection in mice and ferrets. In this study, vaccination of
epitope from respiratory syncytial virus, structural vaccinology mice and ferrets elicited broadly cross-reactive antibodies
enabled to generate small, thermally and conformationally that completely protected mice and partially protected ferrets
stable protein scaffolds that accurately mimic the viral epitope against lethal heterosubtypic H5N1 influenza virus challenge
structure and induce neutralizing antibodies [124]. despite the absence of detectable H5N1 neutralizing activity
in vitro. Protection against H5N1 challenge indicates that
5.3. Vaccines pipeline vaccine-elicited hemagglutinin stem-specific antibodies can
Research on molecular biology methods in vaccine develop- protect against diverse group 1 influenza strains [131].
ment has discovered several promising results. There are a
number of novel vaccines at various stages of the drug accep- 5.3.5. Respiratory syncytial virus and Group B
tance process. They include vaccines against major infectious Streptococcus
diseases such as HIV and tuberculosis, a universal flu vaccine, There is a public health need for vaccines against respiratory
and vaccines against noninfectious diseases. syncytial virus and Group B Streptococcus. For both pathogens,

Biotechnology and Applied Biochemistry 315

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maternal immunization could reduce the risk of neonatal in- to the expected growth of the vaccines market include high
fection and death by passive placental transfer of maternal prevalence of diseases, rising government and nongovernment
antibodies. New vaccines against these conditions may es- funding for vaccine development, and increasing focus on
tablish a precedent for maternal immunization as the initial immunization programs [137].
indication [132]. Respiratory syncytial virus is a pathogen for For example, market for vaccines against shingles is
which there is significant clinical pipeline activity, and the predicted to more than double by 2022 [138]. Currently,
likelihood of a candidate emerging for licensure soon. One Zostavax, live, attenuated virus vaccine, dominates this market.
company is considering phase III study designs for their Group However, it is anticipated that Shingrix, a recombinant subunit
B Streptococcus candidate vaccine [132]. vaccine, which is under review by regulators in the United
States and Europe, may offer greater protection in older
5.4. Noninfectious disease vaccines patients [138].
As life expectancy has increased in the recent past, nonin- The global vaccines market is segmented based on tech-
fectious diseases such as ischemic heart disease, stroke, and nology, type, disease indication, end-users, and regions. Based
cancer are the leading cause of death. Some other noninfectious on increasing company investments, the highest growth rate
diseases, including diabetes, Alzheimer’s disease, and other in the vaccines market is expected to be registered in the
neurodegenerative diseases, are becoming leading causes of conjugate vaccines segment. Two conjugate vaccines against
morbidity [133]. The development of therapeutic vaccines will Streptococcus pneumoniae, Penavnar and Prevnar 13, already
lead to opportunities to manage these diseases at early stages. succeeded in the marked. The combined sales of these vaccines
Vaccines to treat chronic diseases would be expected to extend accounted for approximately $6,3 billion in 2015 which places
life expectancy. Currently, studies of this type are in the discov- them on the top of best-selling vaccines list (Fig. 2).
ery stage. However, in 2010, the FDA approved Sipuleucel-T, In 2016, biopharmaceutical sales (excluding vaccines)
the first therapeutic vaccine that represents a milestone and reached U.S.$163 billion, a 5.8% increase since 2015 and a
may pave the way for the wider use of cancer vaccine im- 102% increase since 2008 [139]. It is believed that this market
munotherapies. Sipuleucel-T is an autologous active cellular has great potential for further dynamic growth. According
immunotherapy used for treating men with asymptomatic or to the report “Global Protein Therapeutics Market Outlook
minimally symptomatic metastatic castration-resistant prostate 2020,” this market may reach U.S.$208 billion by the end of
cancer [134]. 2020 [140]. The increase is a result of the growing number of
Effective therapeutic cancer vaccines may rely on the innovative biopharmaceuticals launched on the market, their
development of personalized vaccines tailored to match a therapeutic efficacy, and the high prices of this group of drugs
person’s particular cancer mutations. Two clinical studies are compared with conventional drugs. Since 1995, approximately
the first to report that the approach could combat melanoma 50 biopharmaceuticals have been registered every 4 years. By
skin cancer in humans [135]. The ideas for these vaccines the end of 2014, a total 212 biopharmaceuticals were registered
are based on using unique cancer cell components that are and approved in the United States and the European Union.
combined with agents that stimulate an immune response. The However, most profits from biopharmaceutical sales are
vaccine is injected into the patient to trigger an immune attack generated by one group of these drugs: monoclonal antibodies.
against cancer.
6.1. Therapeutic antibodies
Since the registration of the first mAb in 1986, the sales of the
6. The Growth of Biopharmaceuticals
mAbs have grown every year and in 2016 it reached U.S.$106.9
Market billion [139]. mAbs sales thus account for nearly 66% of the
Currently, the industry consisting of the development, manufac- total sales of biopharmaceuticals (excluding vaccines) (Fig. 3).
turing, and marketing of biopharmaceuticals is a multibillion This corresponds to a 205% increase since 2008, when the
dollar industry. The common practice in market reports is sales of these preparations amounted to nearly U.S.$35 billion.
to separately present information about vaccines and other For comparison, the sales of all biopharmaceuticals increased
biopharmaceuticals. by 102% in this period.
Vaccine research costs continue to grow. One of the major The top 10 best-selling biopharmaceuticals in 2016 in-
contributing factors to this growth is the use of state-of-the-art cluded eight Abs (six mAbs and two Fc-based fusion proteins)
vaccine development techniques. On the other hand, society (Fig. 4). The monoclonal antibody adalimumab (brand name
expectation is that a vaccine must be affordable for everyone. Humira), a TNF-α inhibitor used to treat rheumatoid arthri-
Because of that implied price cap, pharma industry generally tis and related disorders, ranked first on this list, generating
regards vaccines as not the most profitable market segment. revenue of $16.486 billion.
However, this perception of vaccine market is changing. In addition to mAbs, the most profitable biopharmaceu-
In 2015, worldwide vaccination market generated 27.6 ticals in the group of the top 10 best-selling products include
billion U.S. dollars and is projected to total around 39.0 billion insulin glargine (long-acting basal insulin analogue) and peg-
U.S. dollars in 2022 [136]. The major factors contributing filgrastim (a factor-stimulating granulocyte colony formation)

316 Progress in Biopharmaceutical Development

 Sales (US$ billion) of the five best-selling vaccines
FIG. 2 in 2015. Prepared based on the data published by
EvaluatePharma 2016 [136].

The shares of recombinant proteins and mAbs in

FIG. 3 the world market of therapeutic proteins sales for development of biosimilars reduces time and cost required
2016. Prepared based on the data published by La (Fig. 5). The benefits associated with the introduction of
Merie in 2017 [139]. biosimilars include reduced therapy costs, increased availabil-
ity of the therapy, and, consequently, more balanced health
(Fig. 3). In 2016 similarly to 2015, these drugs generated total care expenditure [4, 141].
profits of U.S.$11.6 billion [139]. However, the total savings resulting from the administra-
tion of biosimilars will not be as significant as that resulting
6.2. Biosimilars from of the replacement of original synthetic drugs with
Therapy using biopharmaceuticals is costly. For example, in generics. This is because the manufacture and introduction of
2009, the annual cost of breast cancer therapy with Herceptin biosimilars require considerable outlays. It is estimated that
was $37,000 in the United States, whereas that of Gaucher dis- the total cost for the development of a biosimilar drug that
ease therapy with Cerezyme was $200,000 [24]. To reduce the meets the formal requirements for its approval, including the
costs of biopharmaceutical treatment, it is crucial to register cost of manufacturing, could be as high as U.S.$75–250 million
biosimilars based on the documentation of their biosimi- [142], and the whole process could take 7–8 years [143]. These
larity. When compared with reference biopharmaceuticals, are barriers against the introduction of biosimilars on the

Biotechnology and Applied Biochemistry 317

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Sales (US$ billion) of the 10 best-selling

FIG. 4 biopharmaceuticals in 2016. Prepared based on
the data published by La Merie in 2017 [139].

Biosimilars and reference biopharmaceuticals 2006–2008. Since then, there has been a decrease in the
FIG. 5 development timeline [142, 143]. number of registered drugs of this group [51]. The lower-
than-expected number of registered preparations may have
resulted, among other factors, from the scope of the data that
market. Additionally, it is difficult for biosimilars to gain access must be presented for biosimilar approval. This scope is closer
to the market. For example, within two years of its launch on to that required for the registration of an innovative drug,
the market, a biosimilar version of erythropoietin gained a rather than a conventional generic. Moreover, in accordance
37% share in the European market. For comparison, a typical with the regulations adopted in most EU countries, pharmacists
generic drug gains a 90% share in the market within 1 year are not allowed to automatically replace biopharmaceuticals
of its launch [144]. Despite these problems, according to the with biosimilars. The reduction in the number of registered
data of the European Generic Medicines Agency, biosimilars biosimilars may also, at least partially, result from distrust on
generated savings of approximately 1.4 billion EUR in the EU the parts of both patients and doctors [149]. Therefore, the
in 2009. The median decrease in the market prices of the success of biosimilars will depend not only on the quality of the
reference biopharmaceuticals caused by the introduction of preparations but also on developing the trust of doctors and
biosimilars amounted to 35% (data for 2006–2013). Moreover, it patients.
is estimated that in 2007–2020, in eight EU countries (France,
Germany, Italy, Poland, Romania, Spain, Sweden, and the 6.3. Biosimilar market forecast
UK), biosimilars of erythropoietins, G-CSFs, and monoclonal According to Allied Market Research, the revenue of the world
antibodies will generate between 11.8 and 33.4 billion EUR in biosimilar market will increase from U.S.$2.55 billion in 2014
savings [145]. to U.S.$26.55 billion in 2020, growing at a CAGR of 49.1%
However, it must be emphasized that the greatest num- from 2015 to 2020 [147]. This increase in the market value
bers of biosimilar registrations in the EU were recorded in will be influenced by the sales of biosimilars, which will likely

318 Progress in Biopharmaceutical Development

occur after the expiration of the patents for the reference Author Contribution
drugs that currently bring the highest profits, among other
Malgorzata Kesik-Brodacka was responsible for the conception
factors.
and development of this review article, critically revised the
content of the manuscript at all stages, and provided final
approval of the submitted version. Malgorzata Kesik-Brodacka
7. Future Prospects for is the guarantor for the overall content.
Biopharmaceuticals
In recent years, the biopharmaceutical market has been Conflict of Interest
developing at a faster rate than the market for all drugs.
MKB is a coinventor of patents and patent applications per-
According to analysts, this market will continue to grow.
taining to insulin analogs, avian influenza flu vaccine, and
The recently observed and anticipated steady increases in
recombinant proteins expression systems.
the sales of biopharmaceuticals are associated, among other
factors, with the growth of the elderly population and the
consequent increase in the number of chronic diseases, the
growing number of diabetes and cancer patients, and an
8. References
[1] Rader, R (2008) Nat. Biotechnol. 26, 743–751.
increase in the incidence of autoimmune diseases. Insight into
[2] Craik, D. J., Fairlie, D. P., Liras, S., and Price, D. (2013) Chem. Biol. Drug Des.
the mechanisms underlying various medical conditions has 81, 136–147.
facilitated the identification of specific factors and processes [3] Mitragotri, S., Burke, P. A., and Langer, R. (2014) Nat. Rev. Drug Discov. 13,
triggering the pathological changes [148]. This has inspired 655–672.
continued research on the applicability of biopharmaceuticals [4] Misra, M. (2012) Indian J. Pharmacol. 44, 12–14.
[5] Weise, M., Kurki, P., Wolff-Holz, E., Bielsky, M. C., and Schneider, C. K. (2014)
in new clinical situations.
Blood 124, 3191–3196.
The confirmed efficacy of biopharmaceutical drugs and [6] Dimitrov, D. S. (2012) Methods Mol. Biol. 899, 1–26.
their acceptance as therapeutic solutions by doctors and [7] Carter, P. J. (2011) Exp. Cell Res. 317, 1261–1269.
patients all contribute to the growing demand for new biophar- [8] Morishita, M., and Peppas, N. A. (2006) Drug Discov. Today. 11, 905–9010.
maceuticals. One advantage of their application is that they [9] Molowa, D. T., and Mazanet, R. (2003) Biotechnol. Annu. Rev. 9, 285–302.
[10] Kessler, M., Goldsmith, D., and Schellekens, H. (2006) Nephrol. Dial.
offer targeted therapies rather than symptomatic treatment
Transplant. 21, v9–v12.
[146]. [11] Sauerborn, M., Brinks, V., Jiskoot, W., and Schellekens, H. (2010) Trends
In many cases, they have facilitated the treatment of Pharmacol. Sci. 31, 53–59.
previously incurable diseases. [12] Guideline on immunogenicity assessment of biotechnology-derived
The ability to produce proteins with properties superior therapeutic proteins. Available at http://www.ema.europa.eu/docs/en_
GB/document_library/Scientific_guideline/2009/09/WC500003946.pdf.
to those of native proteins has played a major role in this
Accessed 25 Feb 2017.
regard. The growth rate of the biopharmaceuticals market [13] Crommelin, D., Bermejo, T., Bissig, M., Damiaans, J., Kramer, I., Rambourg,
may be significantly influenced by the development of molec- P., Scroccaro, G., Strukelj, B., and Tredree, R. (2005) Eur. J. Hosp. Pharm.
ular biology methods and their automation, increases in the Sci. 11, 11–17.
knowledge about expression systems, and better understand- [14] Schellekens, H., and Casadevall, N. (2004) J Neurol. 251, II/4-II/9.
[15] Wroblewski, M. S., Jex, E. A., Drennon, Munck S., Garmon, C.
ing of the operational processes and technological factors
J., Michel, S. T., Wantman, A., and Neal, O. L. (2009) Emerging
related to the scale-up of recombinant protein production. The health care issues: follow-on biologic drug competition. Available at
highly promising prospects of the biopharmaceutical market http://www.ftc.gov/sites/default/files/documents/reports/emerging-health-
are related to breakthrough innovations, such as the develop- care-issues-follow-biologic-drug-competition-federal-trade-commission-
ment of immunotherapy, antibody–drug conjugates, and gene report/p083901biologicsreport.pdf. Accessed 25 Feb 2017.
[16] de Joncheere, K., Rietveld, A. H., and Huttin, C. (2002) Int. J. Risk Saf. Med.
therapies.
15, 101–109.
Factors hindering the development of this market include [17] Guideline on similar biological medicinal products. Available at
the high costs of implementing the developed biopharmaceuti- http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_gui
cals. deline/2014/10/WC500176768.pdf. Accessed 25 Feb 2017.
In the area of novel biopharmaceuticals, we may see a [18] Weise, M., Bielsky, M. C., de Smet, K., Ehmann, F., Ekman, N., Narayanan,
G., Heim, H. K., Heinonen, E., Ho, K., Thorpe, R., Vleminckx, C., Wadhwa,
trend in the active pharmaceutical ingredients toward the
M., and Schneider, C. K. (2011) Nat. Biotechnol. 29: 690–693.
enhancement of naturally found ones’ therapeutic efficacies. [19] Cai, X. Y., Wake, A., and Gouty, D. (2013) Bioanalysis 5, 517–520.
Considering the data on the preparations being currently tested [20] Scientific Considerations in Demonstrating Biosimilarity to a Refer-
in clinical trials, we can expect a steady increase in the numbers ence Product. U.S. Department of Health and Human Services Food
of newly registered mAbs and their dominant presence in the and Drug Administration. Available at https://www.fda.gov/downloads/
Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM29
biopharmaceutical market. Because the patent protections of
1128.pdf. Accessed 17 Oct 2017.
many of the best-selling biopharmaceuticals will expire soon, [21] Vlasak, J., Bussat, M. C., Wang, S., Wagner-Rousset, E., Schaefer, M.,
it appears logical to forecast the introduction of a significant Klinguer-Hamour, C., Kirchmeier, M., Corvaı̈a, N., Ionescu, R., and Beck, A.
number of biosimilars, which will be their equivalents. (2009) Anal. Biochem. 392, 145–154.

Biotechnology and Applied Biochemistry 319

 Biotechnology and
Applied Biochemistry

[22] Khawli, L. A., Goswami, S., Hutchinson, R., Kwong, Z. W., Yang, J., Wang, [49] Sanchez-Garcia, L., Martin, L., Mangues, R., Ferrer-Miralles, N., Vázquez, E.,
X., Yao, Z., Sreedhara, A., Cano, T., Tesar, D., Nijem, I., Allison, D. E., Wong, and Villaverde, A. (2016) Microb. Cell Fact. 15, 33.
P. Y., Kao, Y. H., Quan, C., Joshi, A., Harris, R. J., and Motchnik, P. (2010) [50] Estes, S., and Melville, M. (2014) Adv. Biochem. Eng. Biotechnol. 139, 11–33.
mAbs. 2, 613–624. [51] Walsh, G. (2014) Nat. Biotechnol. 32, 992–1000.
[23] Beck, A. (2011) mAbs. 3, 107–110. [52] Huang, X., Wang, X., Zhang, J., Xia N., and Zhao, Q. (2017) NPJ Vac. 2, 3.
[24] Johnson, J. A. FDA regulation of follow-on biologics. Avail- [53] Kesik-Brodacka, M., Romanik, A., Mikiewicz-Sygula, D., Plucienniczak, G.,
able at https://primaryimmune.org/wp-content/uploads/2014/05/ and Plucienniczak, A. (2012) Microb. Cell Fact. 16, 109.
Biosimilars_Congressional_Research_Service_Report.pdf. Accessed 17 [54] Baeshen, M. N., Al-Hejin, A. M., Bora, R. S., Ahmed, M. M. M., Ramadan,
Oct 2017. H. A. I., Saini, K. S., Baeshen, N. A., and Redwan, E. M. (2015) J. Microbiol.
[25] European public assessment reports. Available at http://www.ema. Biotechnol. 25, 953–962.
europa.eu/ema/index.jsp?curl=pages%2Fmedicines%2Flanding%2Fe [55] Zeltins, A. (2013) Mol. Biotechnol. 53, 92–107.
par_search.jsp&mid=WC0b01ac058001d124&searchTab=searchByAuth [56] Gupta, S. K., and Shukla, P. (2017) Front. Pharmacol. 8, 419.
Type&alreadyLoaded=true&isNewQuery=true&status=Authorised&key [57] Dalton, A. C., and Barton, W. A. (2014) Protein Sci. 23, 517–525.
word=Enter±keywords&searchType=name&taxonomyPath=&treeNumber [58] Darby, R. A., Cartwright, S. P., Dilworth, M. V., and Bill, R. M. (2012) Methods
=&searchGenericType=biosimilars&genericsKeywordSearch=Submit. Mol. Biol. 866, 11–23.
Accessed 23 Feb 2017. [59] Ahmad, M., Hirz, M., Pichler, H., and Schwab H. (2014) Appl. Microbiol.
[26] Schneider, C. K., Vleminckx, C., Gravanis, I., Ehmann, F., Trouvin, J. Biotechnol. 98 5301–5317.
H., Weise, M., and Thirstrup, S. (2012) Nat. Biotechnol. 30, 1179– [60] Sivakumar G., Ed. (2017) New Insights into Cell Culture Technology.
1185. pp. 43–97, InTech, Rijeka, Croatia.
[27] FDA approves first biosimilar product Zarxio. Available at http://www. [61] Rohricht, P. (1999) Biopharm.-Appl. T. Biol. 12, 46–49.
fda.gov/NewsEvents/Newsroom/PressAnnouncements/ucm436648.htm. [62] Rohricht, P. (1999) Biopharm.-Appl. T. Biol. 12, 52–54.
Accessed 23 Feb 2017. [63] Bertolini, L. R., Meade, H., Lazzarotto, C. R., Martins, L. T., Tavares,
[28] Strohl, W. R. (2015) BioDrugs 29, 215–239. K. C., Bertolini, M., and Murray, J. D. (2016) Transgenic Res. 25, 329–
[29] Zalevsky, J., Chamberlain, A. K., Horton, H. M., Karki, S., Leung, I. W., 343.
Sproule, T. J., Lazar, G. A., Roopenian, D. C., and Desjarlais, J. R. (2010) Nat. [64] Summary basis for regulatory action-ATryn. Available at https://way
Biotechnol. 28, 157–159. back.archive-it.org/7993/20170406141638/https://www.fda.gov/Biologics
[30] LoRusso, P. M., Weiss, D., Guardino, E., Girish, S., and Sliwkowski, M. X. BloodVaccines/BloodBloodProducts/ApprovedProducts/Licensed
(2011) Clin. Cancer Res. 17, 6437–6447. ProductsBLAs/FractionatedPlasmaProducts/ucm134048.htm. Accessed
[31] Verma, S., Miles, D., Giann, L., Krop, I. E., Welslau, M., Baselga, J., Pegram, 17 Oct 2017.
M., Oh, D-Y., Diéras, V., Guardino, E., Fang, L., Lu, M. W., Olsen, S., and [65] FDA approves new product to treat rare genetic disease. Available at
Blackwell, K. (2012) N. Engl. J. Med. 367, 1783–1791. https://wayback.archive-it.org/7993/20170112211438/http://www.fda.
[32] Anou, R. (2014) GaBI J. 3, 166–167. gov/NewsEvents/Newsroom/PressAnnouncements/ucm405526.htm.
[33] Di Nardo, G., and Gilardi, G. (2012) Int. J. Mol. Sci. 13, 15901–15924. Accessed 17 Oct 2017.
[34] Ortiz de Montellano, P. R., Ed. (2005) Cytochrome P450: Structure, Mecha- [66] Moura, R. R., Melo, L. M., and de Figueirêdo Freitas, V. J. (2011) Braz. Arch.
nism, and Biochemistry. pp. 183–246. Kluwer Academic Publishers, New boil. Technol. 54, 927–938.
York. [67] Guideline on the quality of biological active substances produced
[35] Denisov, G., Makris, T. M., Sligar, S. G., and Schlichting, I. (2005) Chem. Rev. by stable transgene expression in higher plants. Available at
105, 2253–2278. http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guide
[36] Di Nardo, G., Fantuzzi, A., Sideri, A., Panicco, P., Sassone, C., Giunta, C., and line/2009/09/WC500003154.pdf. Accessed 17 Oct 2017.
Gilardi, G. (2007) J. Biol. Inorg. Chem. 12, 313–323. [68] Kesik-Brodacka, M., Lipiec, A., Kozak Ljunggren, M., Jedlina, L., Miedzinska,
[37] Tsotsou, G. E., Di Nardo, G., Sadeghi, S. J., Fruttero, R., Lazzarato, L., K., Mikolajczak, M., Plucienniczak, A., Legocki, A.B., and Wedrychowicz, H.
Bertinaria, M., and Gilardi, G. (2013) J. Biomol. Screen. 18, 211–218. (2017) PLoS Negl. Trop. Dis. 11, e0005451.
[38] Tsotsou, G. E., Sideri, A., Goyal, A., Di Nardo, G., and Gilardi, G. (2012) [69] Lomonossoff, G. P., and D’Aoust, M. A. (2016) Science. 16, 1237–1240.
Chemistry 18, 3582–3588. [70] Yao, J., Weng, Y., Dickey, A., and Wang K. Y. (2015) Int. J. Mol. Sci. 16,
[39] Di Nardo, G., Dell’Angelo, V., Catucci, G., Sadeghi, S. J., and Gilardi, G. 28549–28565.
(2015) Arch. Biochem. Biophys. S0003–S9861, 30116–30118. [71] Tekoah, Y., Shulman, A., Kizhner, T., Ruderfer, I., Fux, L., Nataf, Y., Bartfeld,
[40] Capoferri, L., Leth, R., TerHaar, E., Mohanty, A. K., Grootenhuis, P. D., D., Ariel, T., Gingis–Velitski, S., Hanania, U. and Shaaltiel, Y. (2015) Plant
Vottero, E., Commandeur, J. N. M., Vermeulen, N. P. E., Jørgensen, F. S., Biotechnol. J. 13, 1199–1208.
Olsen, L., and Geerke D. P. (2016) Proteins 84, 383–396. [72] Fox, J. L. (2012) Nat. Biotechnol. 30, 472.
[41] Dodhia, V. R., Fantuzzi, A., and Gilardi, G. (2006) J. Biol. Inorg. Chem. 11, [73] FDA approves new orphan drug to treat a form of Gaucher dis-
903–916. ease. Available at https://wayback.archive-it.org/7993/20161022200206/
[42] Castrignanò, S., Ortolani, A., Sadeghi, S. J., Di Nardo, G., Allegra, P., and http://www.fda.gov/NewsEvents/Newsroom/PressAnnouncements/ucm
Gilardi, G. (2014) Anal. Chem. 86, 2760–2766. 302549.htm. Accessed 17 Oct 2017.
[43] Rua, F., Sadeghi, S. J., Castrignanò, S., Di Nardo, G., and Gilardi, G. (2012) [74] Zemella, A., Thoring, L., Hoffmeister, C., Kubick, S. (2015) Chembiochem.
J. Inorg. Biochem. 117, 277–284. 16, 2420–2431.
[44] Sadeghi, S. J., and Gilardi, G. (2013) Biotechnol. Appl. Biochem. 60, 102–110. [75] Oza, J. P., Aerni, H. R., Pirman, N. L., Barber, K. W., ter Haar, C. M., Rogulina,
[45] Rua, F., Sadeghi, S. J., Silvia Castrignanò, S., Valetti, F., and Gilardi, G. (2015) S., Amrofell, M. B., Isaacs, F. J., Rinehart, J., and Jewett M. C. (2015) Nat.
Bioelectrochemtry 105, 110–116. Commun. 6, 8168.
[46] Degregorio, D., D’Avino, S., Castrignanò, S., Di Nardo, G., Sadeghi, S. J., [76] (a) Shinoda, T., Shinya, N., Ito, K., Ishizuka-Katsura, Y., Ohsawa, N., Terada, T.,
Catucci, G., and Gilardi, G. (2017) Front. Pharmacol. 8, 121. Hirata, K., Kawano, Y., Yamamoto, M., Tomita, T., Ishibashi, Y., Hirabayashi,
[47] Zawada, J. F., Yin, G., Steiner, A. R., Yang, J., Naresh, A., Roy, S. M., Gold, Y., Kimura-Someya, T., Shirouzu, M., and Yokoyama, S. (2016) Sci. Rep. 6,
D. S., Heinsohn, H. G., and Murray, C. J. (2011) Biotechnol. Bioeng. 108, 30442; (b) Yuan, L. (2017) Synth. Syst. Biotechnol. 2, 23–27.
1570–1578. [77] Li, X., Abrahams, C., Embry, M., Yu, A., Kahana, J., Brown, M., Narla, R. K.,
[48] Dumont, J., Euwart, D., Mei, B., Estes, S., and Kshirsagar, R. (2016) Crit. Rev. Barnes, L., Schwartz, E., Boylan, J., Zawada, J., Stephenson, H., Bruhns, M.,
Biotechnol. 36, 1110–1122. Bussell, S., Steiner, A., Galan, A., Kline, T., Yam, A., Stafford, R., Hoffmann,

320 Progress in Biopharmaceutical Development

 H., Matheny, S., DeAlmeida, V., Vasquez, N., Heinsohn, H., Sato, A., Molina, Anand, B., Morrison, K., Pereira, D. S., and Stover, D. (2014) PLoS One 9,
A., Hallam, T., and Lupher, M. (2016) Blood. 128, 464. e83865.
[78] Moran, N. (2012) Nat. Biotechnol. 30, 1153. [111] FDA approves Gazyva for chronic lymphocytic leukemia. Available at https://
[79] Whalen, J. (2012) Wall Street J. Available at http://www.wsj.com/ wayback.archive-it.org/7993/20161022101354/http://www.fda.gov/News
articles/SB10001424052970203707604578095091940871524. Events/Newsroom/PressAnnouncements/ucm373209.htm. Accessed 17 Oct
[80] Morrison, C. (2015) TradeSecrets. Available at http://blogs.nature.com/ 2017.
tradesecrets/2015/03/03/1-million-price-tag-set-for-glybera-gene-therapy. [112] Mössner, E., Brünker, P., Moser, S., Püntener, U., Schmidt, C., Herter,
[81] Touchot, N., and Flume, M. (2017) Genes. 8, 78. S., Grau, R., Gerdes, C., Nopora, A., van Puijenbroek, E., Ferrara, C.,
[82] Pearson, S., Jia, H., and Kandachi, K. (2004) Nat Biotechnol. 22, 3–4. Sondermann, P., Jäger, C., Strein, P., Fertig, G., Friess, T., Schüll, C., Bauer,
[83] FDA approves first-of-its-kind product for the treat- S., Dal Porto, J., Del Nagro, C., Dabbagh, K., Dyer, M. J. S., Poppema, S.,
ment of melanoma. Available at https://www.fda.gov/News Klein, C., and Umaña, P. (2010) Blood 115, 4393–4402.
Events/Newsroom/PressAnnouncements/ucm469571.htm [113] Cornes, P. (2012) Targ. Oncol. 7, S57-S67.
[84] Summary of opinion (initial authorisation). Available at http://www. [114] Reichert, J. (2017) New antibodies for cancer start clinical testing. Available
ema.europa.eu/docs/en_GB/document_library/Summary_of_opinion_- at http://www.antibodysociety.org/2017/04. Accessed 17 Oct 2017/
_Initial_authorisation/human/003854/WC500203918.pdf. [115] Reichert, J. (2017) Update on antibody therapeutics in late-stage clinical
[85] Aiuti, A., Roncarolo, M. G., and Naldini, L. (2017) EMBO Mol. Med. 9, studies. Available at http://www.antibodysociety.org/category/phase-3-
737–740. pipeline/. Accessed 18 Sept 2017.
[86] Gene Therapy Clinical Trials Worldwide Database. J Gene Med. Available at [116] Foged, C. (2011) Ther. Deliv. 2, 1057–1077.
http://www.abedia.com/wiley/ [117] Dowling, J. K., and Mansell, A. (2016) Clin. Transl. Immunol. 5, e85.
[87] Novartis CAR-T cell therapy CTL019 unanimously (10-0) recommended for [118] Didierlaurent, A. M., Laupèze, B., Di Pasquale, A., Hergli, N., Collignon, C.,
approval by FDA advisory committee to treat pediatric, young adult r/r B-cell and Garçon N. (2016) Exp. Rev Vacc. 16, 55–63.
ALL. Available at https://www.novartis.com/news/media-releases/novartis- [119] Modjarrad, K., Koff, W.C., Eds. (2017) Human Vaccines Emerging Tech-
car-t-cell-therapy-ctl019-unanimously-10-0-recommended-approval-fda nologies in Design and Development. pp. 65–86. Academic Press, New
[88] George, L.A., Sullivan, S.K., Giermasz, A., Ducore, J. M., Teitel, J. M., Cuker, York.
A., Sullivan, L. M., Majumdar, S., McGuinn, C. E., Galvao, A. M., Luk, A. Y., [120] Pizza, M., Scarlato, V., Masignani, V., Giuliani, M. M., Aricò, B., Comanducci,
Wright, J. F., Chen, Y., Hui, D. J., Wachtel, K., Urich, T., Takefman, D., Couto, M., Jennings, G. T., Baldi, L., Bartolini, E., Capecchi, B., Galeotti, C. L.,
L. B., Carr, M. E., Anguela, X. M., and High K. A. (2016) Blood. 128, 3. Luzzi, E., Manetti, R., Marchetti, E., Mora, M., Nuti, S., Ratti, G., Santini, L.,
[89] Boudes, P. F. (2014) Eur. J. Intern. Med. 25, 31–36. Savino, S., Scarselli, M., Storni, E., Zuo, P., Broeker, M., Hundt, E., Knapp,
[90] Erdmann, V. A., Barciszewski, J., Eds. (2012) From Nucleic Acids Sequences B., Blair, E., Mason, T., Tettelin, H., Hood, D. W., Jeffries, A. C., Saunders, N.
to Molecular Medicine, Springer-Verlag, Berlin Heidelberg. pp. 19–45. J., Granoff, D. M., Venter, J. C., Moxon, E. R., Grandi, G., and Rappuoli, R.
[91] Barrangou, R., and Doudna, J.A. (2016) Nat. Biotechnol. 34, 933–941. (2000) Science 287, 1816–1820.
[92] Yin, H., Xue, W., Chen, S., Bogorad, R. L., Benedetti, E., Grompe, M., [121] Rappuoli, R., Pizza, M., Del Giudice, G., and De Gregorio E. (2014) Proc. Natl.
Koteliansky, V., Sharp, P. A., Jacks, T., and Anderson, D. G. (2014) Nat. Acad. Sci. U. S. A. 111, 12288–12293.
Biotechnol. 32, 551–553. [122] Dormitzer, P. R., Suphaphiphat, P., Gibson, D. G., Wentworth, D. E.,
[93] Cyranoski D. (2016) Nature 539, 479. Stockwell, T. B., Algire, M. A., Alperovich, N., Barro, M., Brown, D. M., Craig,
[94] Ecker, D. M., Jones, S. D., and Levine, H. L. (2015) mAbs. 7, 9–14. S., Dattilo, B. M., Denisova, E. A., DeSouza, I., Eickmann, M., Dugan, V.
[95] Loertscher, R. (2002) Transplant Proc. 34, 797–800. G., Ferrari, A. R., Gomila, C., Han, L., Judge, C., Mane, S., Matrosovich,
[96] CenterWatch. FDA drug information. Available at http://www. M., Merryman, C., Palladino, G., Palmer, G. A., Spencer, T., Strecker, T.,
centerwatch.com/drug-information/fda-approved-drugs/. Accessed 7 Mar Trusheim, H., Uhlendorff, J., Wen, Y., Yee, A. C., Zaveri, J., Zhou, B., Becker,
2017. S., Donabedian, A., Mason, P. W., Glass, I. J., Rappuoli, R., and Venter, J. C.
[97] Kontermann, R. E. (2012) mAbs. 4, 182–197. (2013) Sci. Transl. Med. 5, 185ra68.
[98] Müller, D., and Kontermann, R. E. (2010) Biodrugs. 24, 89–98. [123] Nuccitelli, A., Cozzi, R., Gourlay, L. J., Donnarumma, D., Necchi, F., Norais,
[99] Chames, P., and Baty, D. (2009) mAbs. 1, 539–547. N., Telford, L. J., Rappuoli, R., Bolognesi, M., Maione, D., Grandi, G., and
[100] Kufer, P., Lutterbüse, R., and Baeuerle, P.A. (2004) Discov Med. 4, 325–332. Rinaudo, C. D. (2011) Proc. Natl. Acad. Sci. U. S. A. 108, 10278–10283.
[101] Müller, D., and Kontermann, R. E. (2007) Curr. Opin. Mol. Ther. 9, 319–326. [124] Correia, B. E., Bates, J. T., Loomis, R. J., Baneyx, G., Carrico, C., Jardine, J.
[102] Kontermann, R. E., and Brinkmann, U. (2015) Drug Discov. Today. 20, G., Rupert, P., Correnti, C., Kalyuzhniy, O., Vittal, V., Connell, M. J., Stevens,
838–847. E., Schroeter, A., Chen, M., MacPherson, S., Serra, A. M., Adachi, Y., Holmes,
[103] Elgundi, Z., Reslan, M., Cruz, E., Sifniotis, V., and Kayser, V. (2016) Adv. Drug M. A., Li, Y., Klevit, R. E., Graham, B. S., Wyatt, R. T., Baker, D., Strong, R. K.,
Deliv. Rev. pii: S0169-409X, 16, 30317–30319. Crowe, J. E., Johnson, P. R., and Schief W. R. (2014) Nature 507, 201–206.
[104] Newman, M. J., and Benani, D. J. (2016) J. Oncol. Pharm. Pract. 22, 639–645. [125] HIV.gov. Available at https://www.niaid.nih.gov/news-events/first-new-hiv-
[105] Bodet-Milin, C., Ferrer, L., Pallardy, A., Eugène, T., Rauscher, A., Faivre- vaccine-efficacy-study-seven-years-has-begun. Accessed 17 Oct 2017.
Chauvet, A., Barbet, J., and Kraeber-Bodéré, F. (2013) Front. Oncol. 3, [126] ClinicalTrials.gov. Available at https://clinicaltrials.gov/ct2/show/NCT0
177. 2788045. Accessed 17 Oct 2017.
[106] Chamarthy, M. R., Williams, S. C., and Moadel, R. M. (2011) Yale J. Biol. [127] Fletcher, H. A., and Schrager, L. (2016) Trans. R. Soc. Trop. Med. Hyg. 110,
Med. 84, 391–407. 212–218.
[107] Wang, Y. J., Li, Y. Y., Liu, X. Y., Lu, X. L., Cao, X., Jiao, B. H. (2017) Mar Drugs. [128] Petersen, E., Maeurer, M., Marais, B., Migliori, G. B., Mwaba, P., Ntoumi, F.,
15, pii: E18. Vilaplana, C., Kim, K., Schito, M., and Zumla, A. (2017) Int. J. Infect. Dis. 56,
[108] Melis, J. (2017) Increased clinical pipeline of antibody drug conjugates in 1–5.
2016. Available at http://www.antibodysociety.org/2017/01. Accessed 12 [129] First malaria vaccine receives positive scientific opinion from EMA.
Aug 2017. Available at http://www.ema.europa.eu/ema/index.jsp?curl=pages/
[109] Sochaj, A. M., Świderska, K. W., and Otlewski, J. (2015) Biotechnol. Adv. 33, news_and_events/news/2015/07/news_detail_002376.jsp&mid=WC0b01ac
775–784. 058004d5c1. Accessed 8 Sept 2017.
[110] Jackson, D., Atkinson, J., Guevara, C. I., Zhang, C., Kery, V., Moon, S. [130] Kesik-Brodacka, M., Plucienniczak, G. (2014) Acta Biochim. Pol. 61, 523–30.
J., Virata, C., Yang, P., Lowe, C., Pinkstaff, J., Cho, H., Knudsen, N., [131] Yassine, H. M., Boyington, J. C., McTamney, P. M., Wei, C. J., Kanekiyo, M.,
Manibusan, A., Tian, F., Sun, Y., Lu, Y., Sellers, A., Jia, X. C., Joseph, I., Kong, W. P., Gallagher, J. R., Wang, L., Zhang, Y., Joyce, M. G., Lingwood,

Biotechnology and Applied Biochemistry 321

 Biotechnology and
Applied Biochemistry

D., Moin, S. M., Andersen, H., Okuno, Y., Rao, S. S., Harris, A. K., Kwong, [138] The commercial outlook for infectious disease vaccines. Available
P. D., Mascola, J. R., Nabel, G. J., and Graham, B. S. (2015) Nat. Med. 21, at https://biopharmadealmakers.nature.com/users/9880-biopharma-
1065–1070. dealmakers/posts/17865-the-commercial-outlook-for-infectious-disease-
[132] Giersing, B. K., Modjarrad, K., Kaslow, D. C., and Moorthy, V. S. (2016) vaccines. Accessed 8 Sept 2017.
Vaccine 34, 2865–2869. [139] 2016 sales of recombinant therapeutic antibodies & proteins. La Merie
[133] Murphy, S. L., Xu, J., and Kochanek, K. D. (2013) Natl. Vital Stat. Rep. 61, Publishing, March 16, 2017.
1–118. [140] Global protein therapeutics market outlook 2020. Available at
[134] Plosker, G. L. (2011) Drugs 71, 101–108. http://www.researchandmarkets.com/reports/2729030/global_protein_
[135] (a) Ott, P. A., Hu, Z., Keskin, D. B., Shukla, S. A., Sun, J., Bozym, D. J., Zhang, therapeutics_market_outlook_2020. Accessed 17 Oct 2017.
W., Luoma, A., Giobbie-Hurder, A., Peter, L., Chen, C., Olive, O., Carter, T. [141] Weise, M., Bielsky, M. C., de Smet, K., Ehmann, F., Ekman, N., Giezen,
A., Li, S., Lieb, D. J., Eisenhaure, T., Gjini, E., Stevens, J., Lane, W. J., Javeri, T. J., Gravanis, I., Heim, H. K., Heinonen, E., Ho, K., Moreau, A.,
I., Nellaiappan, K., Salazar, A. M., Daley, H., Saman, M., Buchbinder, E.I., Narayanan, G., Kruse, N. A., Reichmann, G., Thorpe, R., van Aerts, L.,
Yoon, C. H., Harden, M., Lennon, N., Gabriel, S., Rodig, S. J., Barouch, D.H., Vleminckx, C., Wadhwa, M., and Schneider, C. K. (2012) Blood 120, 5111–
Aster, J. C., Getz, G., Wucherpfennig, K., Neuberg, D., Ritz, J., Lander, E. 5117.
S., Fritsch, E. F., Hacohen, N., and Wu, C. J. (2017) Nature. 547, 217–221; [142] Calo-Fernández, B., and Martı́nez-Hurtado, J.L. (2012) Pharmaceuticals 5,
(b) Sahin, U., Derhovanessian, E., Miller, M., Kloke, B. P., Simon, P., Löwer, 1393–1408.
M., Bukur, V., Tadmor, A.D., Luxemburger, U., Schrörs, B., Omokoko, T., [143] Blackstone, E. A., and Fuhr, J. P. (2013) Am. Health Drug Benefits. 6, 469–478.
Vormehr, M., Albrecht, C., Paruzynski, A., Kuhn, A. N., Buck, J., Heesch, S., [144] Lofton, C. J. (2011) Pharm. Today, 17 67–76.
Schreeb, K.H., Müller, F., Ortseifer, I., Vogler, I., Godehardt, E., Attig, S., Rae, [145] Haustein, R., de Millas, C., Höer, A., and Häussler, B. (2012) GaBI J. 1,
R., Breitkreuz, A., Tolliver, C., Suchan, M., Martic, G., Hohberger, A., Sorn, 120–126.
P., Diekmann, J., Ciesla, J., Waksmann, O., Brück, A. K., Witt, M., Zillgen, [146] Minghetti, P., Rocco, P., Cilurzo, F., Vecchio, L. D., and Locatelli, F. (2012) Drug
M., Rothermel, A., Kasemann, B., Langer, D., Bolte, S., Diken, M., Kreiter, Discov. 17, 63–70.
S., Nemecek, R., Gebhardt, C., Grabbe, S., Höller, C., Utikal, J., Huber, C., [147] Deshmukh, R.World biosimilars/follow-on-biologics
Loquai, C., and Türeci, Ö. (2017) Nature 547, 222–226. market−opportunities and forecasts, 2014–2020. Available at
[136] Evaluate Ltd. 2016. EvaluatePharmaWorld Preview 2016, Outlook https://www.alliedmarketresearch.com/global-biosimilars-market.
to 2022. Available at http://info.evaluategroup.com/rs/607-YGS- Accessed 8 Sept 2017.
364/images/wp16.pdf. Accessed 17 Oct 2017. [148] Bilello, J. A. (2005) Curr. Mol. Med. 5, 39–52.
[137] Bloom, B. R., Lambert, P. H., Eds. (2016) The vaccine book, 2nd ed., Academic [149] Krishnan, A., Mody, R., and Malhotra, H. (2015) Biosimilars 2015, 19–
Press, Elsevier, London. 32.

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