ANTIGEN AND

ANTIBODY
Presented By

DR. DHIREN B. BHOI

DEPT. OF GYNAECOLOGY AND OBSTETRICS
COLLEGE OF VETERINARY SCI. & ANIMAL HUSBANDRY
S. D. AGRICULTURAL UNIVERSITY
GUJARAT (INDIA)
E-mail:- drdhirenvet@gmail.com
Antigen
“Antigens are substances that induce a specific immune
response and subsequently react with the products of a specific
immune response.”

• The word, antigen, originated from the notion that they

can stimulate antibody generation. We now know that the
immune system does not only consist of antibodies.
• An antigen is a molecule that stimulates an immune
response.
• The modern definition encompasses all substances that
can be recognized by the adaptive immune system
(i.e. T cell & B cell).
Characteristics of Antigen
1.IMMUNOGENICITY
The capacity to stimulate the production of

antibodies or cell-mediated immune responses.

2.ANTIGENICITY
Ability to bind antibody

3. COMPLETE ANTIGEN
Antigen has both immunogenicity and antigenicity.
4.INCOMPLETE ANTIGEN
 Also known as hapten
Antigen has only antigenicity.
Incomplete antigens have antigenic determinants,
 They cannot induce immune responses

one example of an incomplete antigen is a hapten, which is

an artificial monovalent epitope)

TWO FUNCTION OF ANTIGEN

1. Immunogenicity
“An ability of antigen which can stimulate the body to
evoke a specific immune response.”
2.Immunoreactivity
“ An ability of antigen which can combine with

corresponding Antibody or sensitized T lymphocyte. “

Structure of Antigen
 Classification
According to their
[1] Exogenous antigens
origin
Exogenous antigens are antigens that have entered the body
from the outside, for example by inhalation, ingestion, or
injection. By endocytosis or phagocytosis, these antigens are
taken into the antigen-presenting cells (APCs) and processed
into fragments. e.g. foreign cells such as bacteria, virus

[2] Endogenous antigens

Endogenous antigens are antigens that have been generated
within the cell, as a result of normal cell metabolism, or because
of viral or intracellular bacterial infection. e.g autologus
antigen or autoantigen
[3] Autoantigens
An autoantigen is usually a normal protein or complex of proteins that
is recognized by the immune system of animals suffering from a specific
autoimmune disease. e.g. rheumatoid arthritis and immune thyroiditis
 [4] Tumor antigens
Tumor antigens are those antigens that are presented by the MHC I
molecules on the surface of tumor cells. These antigens can sometimes
be presented only by tumor cells and never by the normal cells. In this
case, they are called tumor-specific antigens (TSAs) and typically
result from a tumor specific mutation.
According to relative
[1] xenoantigen
The prefix "xeno" means foreign or other. It comes from the Greek
"xenos" meaning stranger, guest, or host.

An antigen that is found in more than one species. An antigen is

something that is capable of inducing an immune response.

[2] Alloantigen

An antigen existing in alternative (allelic) forms in a species, thus

inducing an immune response when one form is transferred to members
of the species who lack it; typical alloantigens are the blood group
antigens.
Other Ag.
[1]Complete antigen
antigens with both immunogenicity and antigenicity
[2]hapten: antigens with antigenicity but not immunogenicity
[3]Thymus depended and independed Antigen

Thymus dependent antigen [TD-Ag]

Require accession of T and APC to active B
cell
initial both humor and cell immunity
induce many types of antibody
induce immune memory
Thymus independent antigen, TI-Ag
Directly active B cell independent T cell
Only initial humor immunity and induce
IgM
not induce immune memory
 TD antigen TI-1 antigen TI-2 antigen

chemical protein lipin Polysaccharides

property polysaccharides , glycans
Epitope type T, B epitope reduplicative B bulk
epitope reduplicative B
epitope
B cell clone Most B2 Mature and Mature B1
activated immature B1,B2

Neogenesis respond respond Non respond

mouse and baby
• Heterophile antigen
 Common antigen owned by different germ lines
 Forssman was the first to find that there were
common antigen between cony pig organ and sheep
erythrocytes , which was called Forssman antigen.

• Idiaotype antigen
 TCR 、 BCR or Ig can induce antibody production
too.
Superantigen (SAg)

 Antigens that can non-specifically stimulate a

plenty of T/B cells and induce a very strong Ir
with a extremely low concentration

 The mechanism of their action is different from

that of common Ags
Heterogenous Ag (xenoantigen)
“An antigen that is found in more than one species.”
(1) Microbial Ag
 Surface antigen [“K” Ag]
Pillus Ag [”F” Ag ]
Somatic Ag
 Flagellar Ag [ “H” Ag]
(2) EXOTOXIN AND TOXOID
EXOTOXIN: Produced by G+ve bacteria
Strong immunogenicity and pathogenicity
TOXOID: Under suitable condition exotoxin loss
its toxicity without affecting it’s
immunogenicity then exotoxin
turned toxoid. e.g. tetanus toxoid
(3) HETEROPHILE ANTIGEN
Common antigen owned by different germ lines
 ANTIGENIC EPITOPES
Also known as Antigenic determinants.

“It is the part of a macromolecule( large molecules) that is

recognized by the immune system, specifically by antibodies, B
cells, or T cells.”
The part of an antibody that recognizes the epitope is called a
paratope.
TWO DIFFERENT EPITOPES

• B cell epitope, a portion • T cell epitope, the

of antigen molecule that region of antigen
is recognized by B cell molecules that are
receptors (BCR). recognized by T cell
receptors (TCR).
 TWO TYPE OF EPITOPE
(A) Linear epitopes
continuous and found in polysaccharides as well as in both
native (nondenatured) and denatured proteins, especially
fibrillar proteins.
specificity depends upon primary sequence.
typical size is 5-6 subunits in length
(B) CONFORMATIONAL EPITOPE

Discontinuous (involve multiple subunits, often located far

apart in the primary sequence of the antigen molecule) and are
thus found only in native (globular) proteins.
 ADJUVANT
“An adjuvant is an agent that may stimulate the immune system
and increase the response to a vaccine, without having any
specific antigenic effect in itself.”
"An immunologic adjuvant is defined as any substance that acts
to accelerate, prolong, or enhance antigen-specific immune
responses when used in combination with specific vaccine
antigens”
CLSSIFICATION OF ADJUVANT
1.Freund’s adjuvant
Complete Freund’s adjuvant (CFA)
 Incomplete Freund’s adjuvant (IFA)
2.Liposome
3.Inorganic compound
There are many adjuvants, some of which are inorganic (such as
alum), that also carry the potential to augment immunogenicity
4.Cytokine
5.Biodegradable nanoparticles
6.Organic adjuvants
organic adjuvants are more commonly used
in animal vaccines.
 Antibody [immunoglobulin]
DEFINITION
Immunoglobulin (Ig)

Immunoglobulin are
glycoprotein
molecules that are
produced by plasma
cells in response to an
immunogen and
protect the body
against infection.
CH : constant region of heavy chain
VH : variable region of light chain
CL : Constant region of light chain
VL : variable region of light chain
Fab: fragment antigen binding
Fc : fragment crystallizable
A. Heavy and Light Chains
All immunoglobulins have a four chain structure as their
basic unit. They are composed of two identical light chains
and two identical heavy chains .
e.g. Ig G =λ chain

B. Variable (V) and Constant (C) Regions

• 1. Light Chain - VL (110 amino acids) and CL (110 amino acids)
• 2. Heavy Chain - (110 amino acids) and CH (330-440 amino
acids)

C. Hinge Region
This is the region at which the arms of the antibody molecule forms
a Y. It is called the hinge region because there is some flexibility in
the molecule at this point.
• D. 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 and by
non-covalent interactions 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.

E. IMMUNOGLOBULIN FRAGMENTS:
(A) Fab
Digestion with papain breaks the immunoglobulin molecule in
the hinge region before the H-H inter-chain disulfide bond.
It has antigenic binding site.
B. Fc
Digestion with papain also produces a fragment that
contains the remainder of the two heavy chains each
containing a CH2 and CH3 domain. This fragment was
called Fc because it was easily crystallized.
They have plecental transfer site and complement
binding site.
 Clssification Of Antibody On The Basis Of
Heavy chains
1. IgG

IgG is the major Ig in serum - 75% of

serum Ig is IgG.
IgG is the major Ig in extra vascular
spaces
Location: Blood, lymph, intestine
Half-life in serum: 23 days
Complement Fixation: Yes
Placental Transfer: Yes
IgG is a good opsonin.
The term opsonin is used to describe
substances that enhance phagocytosis.
Mol. Wt. : 1,50,000
e.g. Ig G =λ heavy chain
 2.IgM
Mol. Wt.: 9,00,000
e.g. µ heavy chain

J Chain : IgM and IgA both contain

J chain encoded by as separate
gene called the joining chain which
is disulphide bonded to the
tailpieces, stabilising the multimer
Structure: Pentamer
Percentage serum antibodies: 5-10%
Location: Blood, lymph, B cell surface (monomer)
Half-life in serum: 5 days

Complement Fixation: Yes

Placental Transfer: No
Known Functions: First antibodies produced during an infection.
Effective against microbes and agglutinating antigens.
IgM is the third most common serum Ig.
 3.IgA

Mol. wt. :3,20,000

e.g. α (Alpha) heavy chain

IgA is the 2nd most common serum Ig.

Structure: Dimer
Percentage serum antibodies: 10-15%
Location: Secretions (tears, saliva, intestine, milk), blood and
lymph.
Half-life in serum: 6 days
Complement Fixation: No
Placental Transfer: No
Known Functions: Localized protection of mucosal surfaces.
Provides immunity to infant digestive tract.
 4.IgD
Structure: Monomer
Percentage serum antibodies: 0.2%
Location: B-cell surface, blood, and

lymph
Half-life in serum: 3 days
Complement Fixation: No
Placental Transfer: No
Known Functions: In serum function
is unknown. On B
cell surface, initiate immune
response. Ig D

IgD is found in low levels in serum;

its role in serum Mol. Wt. : 1,80,000
uncertain.
e.g. δ (Delta) heavy chain
 5.IgE
Structure: Monomer
Percentage serum antibodies: 0.002%
Location: Bound to mast cells and
basophils throughout body. Blood.
Half-life in serum: 2 days
Complement Fixation: No
Placental Transfer: No
Known Functions: Allergic reactions.
Possibly lysis of worms.
IgE is the least common serum Ig .
IgE also plays a role in parasitic helminth
diseases.
IgE

Mol. Wt. : 1,90,000

e.g. ε (Epsilon) Heavy chain
 Monoclonal Antibodies

Monoclonal Antibodies - identical antibodies produced in large

quantities by an immortalized hybridoma cell line

Technique developed by Köhler and Milstein in 1975, earned Nobel

Prize in 1984

Benefits: identical antibodies available in unlimited quantity; easily

purified

Drawbacks: may not be useful for all techniques; limited number of

species (mouse, rat, hamster, rabbit)
Monoclonal Antibody Production
•Process by which large quantities of antibodies
(targeted against a particular antigen X) can be
produced.
•A mouse is immunized by injection of an antigen X to stimulate
the production of antibodies targeted against X. The antibody
forming cells are isolated from the mouse's spleen.

•Monoclonal antibodies are produced by fusing single antibody-

forming cells to tumor cells grown in culture. The resulting cell is
called a hybridoma.
•Each hybridoma produces relatively large quantities of
identical antibody molecules. By allowing the hybridoma to
multiply in culture, it is possible to produce a population of
cells, each of which produces identical antibody molecules.
These antibodies are called "monoclonal antibodies"
because they are produced by the identical offspring of a
single, cloned antibody producing cell.
Polyclonal antibody

Polyclonal antibodies are antibodies that are derived from

different B cell lines. They are a mixture of immunoglobulin
molecules secreted against a specific antigen, each recognizing
a different epitope

These antibodies are typically produced by immunization of a

suitable mammal, such as a mouse, rabbit or goat

An antigen is injected into the mammal. This induces the B-

lymphocytes to produce IgG immunoglobulins specific for the
antigen. This polyclonal IgG is polyclonal purified from the
mammal’s serum.
 ANTIGENIC CLASSIFICATION OF ANTIBODY
There are three type: (1) ISOTYPE (2) ALLOTYPE (3) IDIOTYPE
(1) ISOTYPE
“Antigenically different immunoglobulin variants
that are common in all individual of given biological
species are called as isotype.”
e.g. Ig G that is subdivided into subclasses
IgG1,IgG2,IgG3,IgG4.
(2) ALLOTYPE
“Determinants on immunoglobulin molecules that
differ among individual of same species have
inherited different allels.”
(3) IDIOTYPE
“If any antigen determinants or an epitope is localized in a
variable domain of the immunoglobulin chain is called as
Idiotype.”
 Major Histocompatibility Complex (MHC)
 The MHC is a closely linked complex of genes that govern
production of the major histocompatibility

 In humans, MHC resides on the short arm of chromosome 6

 Three genes (HLA-A, HLA-B, HLA-C) code for

 the class I MHC proteins

 Several HLA-D loci determine the class II MHC proteins i.e. DP, DQ
and DR

 HLA genes are very diverse (polymorphic)

 i.e. there are many alleles of the class I and II genes
 Major Histocompatibility Complex (MHC)
 Between the class I and class II gene loci, there is a
third locus (Class III)

 This locus contains genes encoding tumor necrosis

factor, lymphotoxin and two complement components
(C2 and C4)

 Class III antigens do not participate in MHC restriction

or graft rejection
 Major Histocompatibility Complex (MHC)
 Class I MHC antigens are : HLA-A, HLA-B and HLA-C

 These antigens are glycoproteins found on surfaces of

all nucleotide human cells and on platelets

 HLA-A contains 24 different antigenic specificities,

 HLA-B contains 52 and HLA-C contains 11

 Class I MHC antigens are involved of MHC restriction of

cell mediated cytotoxicity
 Major Histocompatibility Complex (MHC)
 Endogenously processed cytosolic peptides in virus
infected cells or tumor cells are transported to the
surface of the cells

 They bind to MHC I molecules to be recognized by

cytotoxic T-cells which then kill these cells

 In other words;
T-cells are only activated when they recognize both
antigen and class I MHC molecules in association
 Major Histocompatibility Complex (MHC)
• Class II antigens are: HLA-DP, HLA-DQ, HLA-
DR antigens

• These antigens are glycoproteins found on the

surface of macrophages, B-cells, Dentritic cells,
langerhans cells of skin and activated T cells

• HLA-DP contain 6 different antigenic specificities,

HLA-DQ contains 9 and HLA-DR contains 20
•
 Class I MHC and Class II MHC

MHC Class I MHC Class II

Nomenclature HLA-A, HLA-B, HLA-C HLA-DP, HLA-DQ,
HLA-DR

Found on All nucleated somatic cells Macrophages, B-cells,

Dentritic cells, langerhans
cells of skin and activated
T cells

Recognized by CD8 TC cells CD4 TH cells

Functions Presentation of Ag to TC Presentation of Ag to TH

cells leading to elimination cells which secrete
of tumor or infected host cytokines
cell
 Types of grafts
• 1) Autografts :
• The transfer of an individual’s own tissues
• from place to place
• e.g. Skin grafts (regularly accepted)

• 2) Isografts :
• Transfer of tissues between genetically
• identical persons
• e.g. Identical twins ( accepted permanently
3) Allografts (homograft):
- Transfer of a graft between genetically different
members of same species
e.g from one human to another
- Rejection occur if donor and recipient are not matched

4) Xenograft (heterograft):
- Transfer of tissues between different species
- Always rejected