Recombinant Insulin

Peter Ko
1101992
Abstract

Recombinant human insulin was historically one of the first products brought forward
from biotechnology and was developed to help combat diabetes. It was manufactured to replace
existing animal insulins that were obtained by modifying pancreatic cells from animals.
Recombinant DNA (rDNA) technologies allowed the production of a wide range of peptides,
proteins, and biochemicals from naturally non producing cells to mass produce insulin to help
meet a growing demand, projected to reach 552 million people by 2030 [7].

Introduction

Diabetes is a disease characterized by genetic, metabolic and autoimmune deficiencies

and abnormalities. Humans need alternatives for people who cannot naturally produce the
protein in their body and need to look towards synthetic alternatives. Companies manufactured
insulin analogues to help with type 1 and type 2 diabetes through recombinant DNA techniques.
Recombinant human insulin was produced via fermentation in (Escherichia coli) for Humulin
and Insumam; Novolin another recombinant form of insulin which was produced in yeast
(Saccharomyces cerevisiae) [1]. With the advent of PCR in the 1970’s it was possible for
researchers to produce millions of copies of specific DNA sequences and pushed the
advancement of genetic engineering.

Human Insulin production

The role of insulin is to manage and control the absorption of glucose from the blood into
the cells where it is utilized as a source of energy and converted into glycogen. It is produced and
stored in the b cells of the pancreas and in the early 80’s was extracted from bovine or porcine
pancreatic tissues[1]. It was later synthesized chemically with recombinant technology which
approached A and B chains and inserted separately of the d-galactosidase structural gene that
controlled the X lac promoter [1]. It was made so that the insulin chains would fuse with the
proteins and join methionine at the end of the P-galactosidase protein which also contained a
Amp’ marker. The transmorats then were screened after plating on a culture medium containing
X-gal and ampicillin. The A and B insulin chains transformants were grown and lysed to harvest
the cells contents. They are then purified separately and because the d-galactosidase gene and the
insulin A gene are fused, the insulin protein produced is a d-galactosidase-insulin hybrid [].

Clinical Studies with Recombinant Human Insulins

The first clinical studies with rInsulin focused on the switch from animal insulin to recombinant
and semisynthetic insulin. These studies included patients with type 1 or 2 diabetes that were
pregnant or paediatric and were studied utilizing a euglycemic clamp technique, a technique
where the plasma insulin concentration is raised and maintained at 100 μU/ml by a continuous
infusion of insulin. This was employed to observe the bioequivalence between the formulas and
the clinical studies were set in ranges of 6 to 12 months and tested for incidences of
hypoglycaemia and levels of antigenicity for insulin-directed antibodies and antibodies to
Escherichia coli or Saccharomyces cerevisiae proteins [3].

The formulations for recombinant human insulin used were soluble regular insulin and
crystalline NPH insulin in premixed ratios. These were found to have similar levels of efficiency
and safety to that of semisynthetic forms of insulin. These studies on bioequivalence also showed
small differences in the time it took for it to take action [3].

Other clinical studies with the recombinant insulin extracted from baker’s yeast
(Novolin) produced similar results in efficiency and safety and was deemed to be a safe and
effective transfer from semi synthetic human insulin to recombinant human insulin without a
change in dosage [].

Studies with forms of recombinant human insulin produced via fermentation in E. coli
showed similar results of proficient levels of bioequivalence and safety when compared with
semi synthetic form of insulin. They were put under extensive clinical studies with type 1 and 2
diabetes who were transferred from semisynthetic to recombinant forms [3].

Therapy Outlook with Recombinant Human Insulin

Recombinant human insulin has several methods of delivery including meal-time

injections which are soluble, basal support, and conventional therapy which is a pre-mix
combination, and disposable or reusable pens.

When a patient switches from one insulin product to another, monitoring is essential even
if it is a similar form of insulin. Initial monitoring of the patient’s glycaemic control should be
unchanged and is particularly important for pre-mixed and NPH-insulins because their
time-action profiles may be different. This is even more essential when a patient exchanges from
a human insulin formulation to an insulin analogue because of an increase in the need for a more
rapid absorption before a meal. This has an insulin profile with a longer duration and requires
fewer injections. Overall changing insulins will change the rate and extent of absorption and will
change the duration of monitoring needed. [3]
Conclusion

Recombinant human insulin revolutionized the way we supply and diagnose patients with
diabetes and it made it possible to mass produce the protein to be more accessible to a growing
population of affected individuals. The application of rInsulin is safe and effective but needs to
be monitored on patients who transition from animal derived insulin or other rInsulin products.
 Bibliography:

1. Landgraf, W., & Sandow, J. (2016, March). Recombinant Human Insulins - Clinical
Efficacy and Safety in Diabetes Therapy. Retrieved from
https://www.ncbi.nlm.nih.gov/pubmed/29632581
2. Mbanya, J. C., Sandow, J., Landgraf, W., & Owens, D. R. (2017, April). Recombinant
Human Insulin in Global Diabetes Management - Focus on Clinical Efficacy. Retrieved
from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5813441/
3. PMC, E. (n.d.). Recombinant Human Insulins - Clinical Efficacy and Safety in Diabetes
Therapy. Retrieved from https://europepmc.org/article/med/29632581
4. Raskin, P., & Clements, R. S. (1991). The use of human insulin derived from baker's
yeast by recombinant DNA technology. Retrieved from
https://www.ncbi.nlm.nih.gov/pubmed/1799914
5. Sandow, J., Landgraf, W., Becker, R., & Seipke, G. (2015, April). Equivalent
Recombinant Human Insulin Preparations and their Place in Therapy. Retrieved from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5819055/
6. Sandow, J., Landgraf, W., Becker, R., & Seipke, G. (2015, April). Equivalent
Recombinant Human Insulin Preparations and their Place in Therapy. Retrieved from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5819055/
7. Zieliński, M., Romanik-Chruścielewska, A., Mikiewicz, D., Łukasiewicz, N.,
Sokołowska, I., Antosik, J., … Płucienniczak, A. (2019, May). Expression and
purification of recombinant human insulin from E. coli 20 strain. Retrieved from
https://www.ncbi.nlm.nih.gov/pubmed/30735706