Antigen and

Antibodies (Immunoglobulis)

Prof. Dr. Batool Hassan Al-Ghurabi

• Antigen (Ag) is substance which when introduced into
the body stimulates the production of an antibody (Ab)
with which it reacts specifically.

• ANTI - GEN: ‘ ANTI’body ‘GEN’erator Substances

that react with antibodies
• IMMUNO – GEN: ‘IMMUN’E response ‘GEN’erator
• Immunogenicity– ability of an Ag to elicit an immune
reaction in the form of a B cell or T cell response
• Antigenicity– just the ability to combine with the
products of the immune responses
• All molecules that are immunogenic are
antigenic

• Antigens that are not immunogenic, but can

take part in immune response
 Classification of Ag
Based on Immunogenicity
1. Complete antigen : Substances which can induce Ab
formation by themselves and can react specifically with
these antibodies

2. Incomplete antigen (haptens): substances unable to

induce Ab formation on its own but can become
immunogenic when linked to proteins, called carrier
proteins. They are of two types: Complex & Simple
 Classification of Ag
Based on origin:
1. Exogenous antigens are antigens that have entered the
body from the outside, for example by inhalation ,
ingestion , or injection . The immune system's response to
exogenous antigens is often subclinical.

2. Endogenous antigens are antigens that have been

generated within previously normal cells as a result of
normal cell metabolism , or because of viral or intracellular
bacterial infection .
 Classification of Ag
• Aut-oantigens is usually a normal protein or complex of
proteins that is recognized by the immune system of
patients suffering from a specific autoimmune disease .
These Ags should, under normal conditions, not be the
target of the immune system, but, due to genetic and
environmental factors, the normal immunological
tolerance for such an Ag has been lost in these patients.
Iso antigens & Heterophile antigen
• Super antigens are a class of antigens that cause
nonspecific activation of T-cells resulting in polyclonal T
cell activation and massive cytokine release. Produced by
» Bacteria » Virus » Mycoplasma. Ex.

1. Staphylococcal enterotoxins 2. Staphylococcal toxic

shock toxin (TSST-1) 3. Streptococcal pyrogenic

4. Retrovirus in mice, is also known to produce

superantigen
ANTIGENIC DETERMINANTS

• These antigens may have several antigenic determinants, called

epitopes, and each epitope can bind with a specific antibody. Thus,
a single antigen can bind to many different antibodies with different
binding sites
Factors of Antigenicity
1. Foreignness: Antigen must be foreignness to immune
system : What substances are foreignness to immune system
? According to Burnnet’s clone selection theory ,
foreignness ( non-self) means substances that never contact
with lymphocytes during embryo period

2. Molecular Size: Larger molecules are highly antigenic

but lower molecular weight are either non antigenic or
weakly
Factors of Antigenicity
- Substances with mol wt > 1,00,000 Da is highly
immunogenic

- Substances with mol wt less than that is weak

immunogenic.

3. Chemical & Structural Complexity : Most naturally

occurring antigens are either proteins or polysaccharides.
Lipids and nucleic acids are less antigenic on their own but
do so when combined with proteins
• In Proteins, the immunogenicity is dependent on its
structural complexity – i.e if the structure is simple (like
chains of single amino acid or simple sugar,) it is less
immunogenic.
‫طالب ارجو تسجيل اسماءكم في سجل الحضور‬
4- Genetic factors
‫ الصف‬of host:
‫تعليقات من‬ 9 Some substances are •
immunogenic in one species or individual but not in other.
The species or individuals may lack or have altered genes
that code for the recognition of antigens.
5- Route of entry, Dose and Physical form Generally, the •
subcutaneous or intramuscular route is better than the
intravenous or intragastric route.

Dose: Very low dose of Ag – do not stimulate immune •

response because too few lymphocytes are contacted.
Increasing the dose or repeating it gives a better immune
response.

Physical form, particulate Ags are more immunogenic than •

soluble and denatured Ag more immunogenic than the
native form
 Antibodies

Prof. Dr. Batool Hassan Al-Ghurabi

Antibody(Ab) or immunoglobulin(Ig) is the large Y
shaped glycoprotein produced by the body’s immune
system when it detects antigens.
They are synthesized by B lymphocytes and secreted by
plasma cells.
Depending on the electrophoretic migration, 3 types of
globulins are present in the blood, namely α, β and γ. So
antibodies are gamma (γ) globulin. .
• Immunoglobulin and antibody are disease-fighting
proteins developed by most vertebrates in response to
a particular antigen.

• Both immunoglobulin and antibody are

glycoproteins.

• Both of them contain similar regions in their

molecules.

• Immunoglobulins are attached to the B

cell membrane while antibodies float in the
circulation.
 Humoral immune response
1. B lymphocytes recognize
specific antigens
– proliferate and
differentiate into
antibody-secreting plasma
cells
2. Antibodies bind to specific
antigens on microbes; destroy
microbes via specific
mechanisms
3. Some B lymphocytes evolve
into the resting state - memory
cells
• Structure of Antibody

• All antibodies have a common basic structure.

A. Each antibody molecule has 4 polypeptide chains.

• Two small identical light chain(L) and two longer

identical heavy chain(H).

• Hence an antibody represented as H2L2.

• Each polypeptide chain contains an (NH3+) amino

terminal and carboxy (COO-) terminal.
Structures of Antibody
B. They are linked by disulfide bonds.

1. Inter-chain disulfide bonds - The heavy and light

chains and the two heavy chains are held together by
inter-chain disulfide bonds.

The number of inter-chain disulfide bonds varies among

different immunoglobulin molecules.
2. Intra-chain disulfide bonds - Within each of the
polypeptide chains there are also intra-chain disulfide
bonds.
C. The central region of heavy chain at which the arms of
the antibody molecule forms a Y is called hinge region
and it is rich in proline.

There is some flexibility in the molecule at this point

D. Variable & constant region

- L & H chains are further divided into variable & constant

region.

-The variable region concern with antigen binding and

constant region concern with various biological functions
like complement activation and binding to cell surface
receptors.
•
E. Domains •
The amino acid sequence in both light and heavy chain is not a •
linear sequence but there are domes and loops due to the
presence of intra chain disulfide bonds these globular areas
called domains.
IMMUNOGLOBULIN FRAGMENTS •

Digestion of Igs with papain breaks the immunoglobulin •

molecule at the hinge region, and produces three fragments, two
identical fragments (Fab) and one fragment (Fc).

A. Fab: Fragment antigen binding - these fragments were called the •

Fab fragments because they contained the antigen binding sites of
the antibody. It is also called paratope, is a part of an antibody
which recognizes and binds to an antigen.
• The combining site of the antibody is created by both VH and VL.
B. Fc : Fragment crystallization- this fragment was called Fc •
because it was easily crystallize
 IMMUNOGLOBULIN CLASSES, SUBCLASSES AND
TYPES
A. Immunoglobulin classes •

The immunoglobulins can be divided into five different •

classes, based on the constant region of the heavy chains.

1. IgG - Gamma heavy chains •

2.IgM - Mu heavy chains •

3.IgA - Alpha heavy chains •

4.IgD - Delta heavy chains •

5.IgE - Epsilon heavy chains •

B. Immunoglobulin Subclasses •
The classes of immunoglobulins can be divided into subclasses •
based on small differences in the constant region of the heavy
chains.
1. IgG Subclasses
a) IgG1 - Gamma 1 heavy chains
•
b) IgG2 - Gamma 2 heavy chains •
•
c) IgG3 - Gamma 3 heavy chains •
•
d) IgG4 - Gamma 4 heavy chains •
2. IgA Subclasses •
a) IgA1 - Alpha 1 heavy chains •
b) IgA2 - Alpha 2 heavy chains •

C. Immunoglobulin Types •
Immunoglobulins can also be classified by the type of light •
chain that they have. Light chain types are based on
differences in the constant region of the light chain.
**Kappa light chains •

**Lambda light chains •

GENERAL FUNCTIONS Of IMMUNOGLOBULINS
A. Primary function: Antigen binding
Immunoglobulins bind specifically to one or a few •
closely related antigens.
B. Secondary function: Effector Functions •
The binding of an antibody to an antigen has no •
direct biological effect. Rather, the significant
biological effects are secondary "effector functions" of
antibodies.
Not every immunoglobulin will mediate all effector •
functions. Such effector functions include:
•
1. Agglutination: Agglutination of particulate •
antigen, including bacteria and viruses. IgM is
particularly suitable for this function. •
2. Opsonization: Opsonization i.e. coating of •
bacteria with antibody's Fab region (IgG). This
facilitates phagocytosis by cells possessing Fc
receptor, e.g. neutrophil, polymorphonuclear
leucocytes".
3. Neutralization: Neutralization of toxins released "
by bacteria e.g. tetanus toxin is neutralized when
specific IgG antibody binds, thus preventing the
toxin binding to it’s receptor. In the case of viruses,
antibodies can hinder their ability to attach to
receptors on host cells.
4. Complement activation (classical pathway):
especially by IgM and IgG, leads eventually to death of
bacteria by the terminal complement components which
make holes in the cell wall, leading to an osmotic death.

5. Precipitation: Precipitation of soluble antigens by

immune complex formation. They can be removed by
phagocytic cells. and can fix complement.
6. Antibody dependent cell mediated
cytotoxicity (ADCC): Antibodies bind to
organisms via their Fab region. Large granular
lymphocytes NK cells, attach via Fc receptors,
and kill these organisms not by phagocytosis
but by release of toxic substances called
perforins.
7. Mucosal protection: This is provided mainly by
IgA. IgA acts chiefly by inhibiting pathogens from
gaining attachment to mucosal surfaces.
 Agglutination
RBC
IgM Antibody RBC RBC

RBC
RBC RBC RBC

RBC
IgG Antibody

RBC RBC

RBC
 Immune
Precipitation

Antigen

Antibody
 IgG
• Monomer, 70-75% of total immuniglobulin
• Secreted in high quantities in secondary exposures
• Cross the placenta
• Most protective because it is capable of carrying out
all effector functions of immunoglobulins.
• Major functions / applications
– neutralize microbes and toxins
– opsonize antigens for phagocytosis
– activate the complement
– protect the newborn
 IgM
• Normally pentamer linked by J chain, but it can also exist
as a monomer on the surface of B cells
• Secreted initially during primary infection
• Cannot cross the placenta
• Major functions / applications
– secreted first during primary exposure
– activates the complement
– functions as Ag receptor on B cell
– the first Ig made by the fetus and the first Ig to be made by a
virgin B cells when it is stimulated by antigen.
– used as a marker of recent infection
 IgA
• Monomeric in serum
• Dimeric with secretory component (J chain) in gastro-
intestinal tract and respiratory tract
• Major function / application
– neutralizes microbes and toxins
• When IgA is found in secretions is also has another
protein associated with it called the secretory piece or
T piece. the secretory piece is made not in plasma cell,
but in epithelial cells and added to the IgA as it passes
into the secretions. The secretory piece helps IgA to be
transported across mucosa and also protects it from
degradation in the secretions.
 IgD
• Monomeric
• Major functions / applications
– present on the surface of B lymphocytes
– functions as membrane receptor
– role unclear
• has a role in antigen stimulated lymphocyte
differentiation
 IgE
• Mediates type I hypersensitivity
• Monomeric
• Major functions / applications
– associated with anaphylaxis
– plays a role in immunity to
helminthic parasites

Serodiagnosis of infectious and non infectious

allergies
(e.g., allergic bronchopulmonary aspergillosis,
parasitic diseases)
Characteristic of Specific Immune response •
1. Discrimination: is the ability to distinguish •
between self and non-self antigens.
2.Specificity: antibodies react specifically with Ag •
that cause their production.
Sometimes Ab can react with similar (not identical) •
Ag, this is called Cross reaction.
3.Clonality: when Ag enter for the first time, just •
those lymphocytes bearing receptors for this Ag are
stimulated then divided after that produce more cells
of the same specificity.
4. Anamnestic response (memory)
I-First exposure (primary humoral immune response)
a- Lag phase: in the first few days, Ab level in serum is zero and no
Ab production.
Lag phase duration depend on, nature of Ag, dose, route of entry
and immune status.
b-Log phase: Ab produce and increasing.
c-Plateau phase or stationary phase: Ab production stop at certain
level.
d-Decline phase: Ab titer decrease because of combination with Ag.
II-Second exposure (secondary humoral immune response)
a-There is memory cells (no need for lag phase).
b-There is high level of Ab.
c- Only those cells with identical receptors will be stimulated (high
affinity).