ANTIBODY

1st year B. Pharm

 Introduction
• Specialized glycoproteins produced by activated B
cells in response to antigen, and is capable of
binding with antigen that triggered its production.

• Serum is subjected to electrophoresis, serum

proteins are separated into 4 fragments-
- albumin
- globulin α, β and γ
• Immunoglobulin (Ig) constitutes 20-25% of total
serum proteins
There are five classes of immunoglobulins-IgG, IgA, IgM, IgD, IgE
 Structure of immunoglobulin
 Y shaped heterodimer
 Consists of four polypeptide chain
 Four polypeptide chain consists of:
i. Heavy chain(H)
ii. Light chain(L)

All four H and L chains are bound to

each other by disulfide bonds and by
non covalent interactions.
There are five classes of H chains- are structurally and antigenically distinct

Immunoglobulin class Heavy chain types

IgG ϒ

IgA α

IgM μ

IgD μ

IgE ε
Light chain are two types;
i. Kappa(κ)
ii. Lambda(λ)

• L chains are composed of 214 amino acids

• whereas number of amino acids in heavy chain varies ranging from 446 (in
α chain) to 576 (in µ chain)
Variable and Constant region
Variable Region
Variable region consists first 110 amino acid in both L and H region.
 Represents antigen binding site of antibody
 Consists of :
i. Hypervariable region
ii. Paratope

Constant Region
• Consists of remaining part of Ig molecule other than variable region.
• Consists of 104 amino acid for light chain
• Consists of 330 to 440 amino acid for heavy chain
Heavy and light chain domains
Heavy and light chains are further
folded into domains
• Light chain contains one variable
domain(VL) and one constant
domain(CH)

• Heavy chains contain one variable

domain(VH) and 3 to 4 numbers of
constant domains(CH)
Hinge region
• Present only in H chain, present between CH1 and CH2
• Consists proline and cysteine
• IgE and IgM do not have hinge region
• Sensitive to enzymatic digestion
Enzymatic digestion
• Papain digestion
• Pepsin digestion
• Mercaptoethanol
 Functions of Immunoglobulins
1) Antigen binding site (by Fab region)
• Bears variable region, involved in interactions with antigen
• Valency of an antibody refers to number of Fab region it possesses.

2) Effector functions (by Fc Region)

• Fixation of complement
Antibody coating the target cell binds to complement through its Fc receptor which
leads to complement mediated lysis of target cell.

• Binding to various cell types

Phagocytes, lymphocytes, mast cells, eosinophils bear Fc receptor (FcR) that bind to
Fc region of immunoglobulin.
 Immunoglobulin G (IgG)

• 70 to 80% of total Ig in the body

• Highest serum concentration
• IgG has four subclasses;
according to amino acid
sequences of constant region of
their γ-heavy chain.
• IgG1, IgG2, IgG3, IgG4
 Functions of IgG

1) Can cross placenta - Provide immunity to fetus and new born.

2) Complement fixing - Fc region of IgG can bind to complement factors.

3) Phagocytosis- IgG1 and IgG3 bind to Fc receptors present on phagocytes.

4) Plays important role in neutralization of toxin

5) Raised after long time following infection and represent chronic or past
infection (recovery)
 Immunoglobulin M (IgM)

• Highest molecular weight (900 kDa)

• Present only in intravascular

compartment

• exists in both monomeric and

pentameric form
 Functions of IgM
1) Acute infection
First antibody to be produced following an infection

2) Complement fixing
Most potent activator of classical complement pathway

3) Fetal immunity
First antibody to be synthesized in fetal life (20 weeks).

4) Protection against intravascular organism

Protection against blood invasion by microorganism.
 Immunoglobulin (IgA)
Second most abundant Ig (10 to
15%)
Exists in both monomeric and
dimeric form
2 types- serum IgA
secretory IgA

a) Serum IgA
- Predominantly in monomeric form
- Interacts with Fc receptors expressed
on immune effector cells
 b) Secretory IgA
Dimeric in nature
Joined by J chain
Another joining segment presesnt k/as secretory
component

Location; milk, saliva, tears, intestinal and

respiratory secretion

Function:
Mediates local or mucosal immunity.
 Subclasses of IgA
Depending upon amino acid sequences in constant region of heavy chain

i. IgA1
Serum IgA compromises 90% IgA1 and 10% IgA2

ii. IgA2
Present in higher concerntration in secretion than serum

IgA2 lacks the disulphide bond between the heavy and light chain
Polysaccharide antigen tend to induce more IgA2 synthesis.
 Immunoglobulin E (IgE)
• Lowest serum concerntration
• Heat liable antibody (inactivated at 56℃ in 1hour)
• Extravascular in distribution

Function
Mediate type I hypersensitivity reaction
Elevated in helminthic infections
 Immunoglobulin D(IgD)

• Resembles IgG structurally

• Found as membrane Ig on the surface of B cells
and acts as B cell receptor along with IgM
• Monoclonal antibodies
Antibodies derived from a single clone of plasma cell, all having same
antigen specificity, produced against single epitope of an antigen

• Polyclonal antibody
When antigen having multiple epitopes enters the body, each epitope
may stimulate one clone of B cells producing one type of antibody.
Resultant antibody mixture is polyclonal antibody