Antigen-Antibody Reaction

Under Supervision of Dr Punit Saraswat HOD,Department of zoology LMC,Jodhpur

Submitted byMonika Prajapat MSc Previous Zoology

What is antigen ?
A substance that enters the body and stimulates the production of an antibody to fight what the immune system perceives as an invader. An antigen is any substance that causes your immune system to produce antibodies against it. An antigen may be a foreign substance from the environment such as chemicals, bacteria, viruses, or pollen. An antigen may also be formed within the body, as with bacterial toxins or tissue cells.

What is antibody ?
Antibodies (also known as immunoglobulins, abbreviated Ig) are gamma globulin proteins that are found in blood or other bodily fluids of vertebrates, and are used by the immune system to identify and neutralize foreign objects, such as bacteria and viruses. They are typically made of basic structural unitseach with two large heavy chains and two small light chainsto form, for example, monomers with one unit, dimers with two units or pentamers with five units.

 Antibodies are produced by a kind of white blood cell called a plasma cell. There are several different types of antibody heavy chains, and several different kinds of antibodies, which are grouped into different ''isotypes'' based on which heavy chain they possess. Five different antibody isotypes are known in mammals, which perform different roles, and help direct the appropriate immune response for each different type of foreign object they encounter.

Antibody Structure
Monomer: A flexible Y-shaped molecule with four protein chains: 2 identical light chains 2 identical heavy chains Variable Regions: Two sections at the end of Ys arms. Contain the antigen binding sites (Fab). Identical on the same antibody, but vary from one antibody to another. Constant Regions: Stem of monomer and lower parts of Y arms. Fc region: Stem of monomer only. Important because they can bind to complement or cells.

Antibody Structure

Immunoglobulin Classes
I. II. III. IV. V. IgG IgM IgA IgD IgE

ANTIGEN-ANTIBODY REACTIONS
The antigens and the antibodies combine specifically with each other. This interaction between them is called Antigen-Antibody reaction. It may be abbreviated as Ag-Ab reaction. These form the basis for humoral immunity or antibody mediated immunity. These reactions form the basis for detection of infectious disease causing agents and also some non-specific Ags like enzymes. When Ag-Ab reactions occur invitro, they are known as serological reactions.

The reactions between Ag and Ab occur in three stages.

1. Primary stage- This reaction involves formation of Ag-Ab complex. The reaction is rapid and it obeys the general laws of physical chemistry and thermodynamics. The two molecules are held together by non-covalent forces, hydrogen bonding, ionic bonding and sometimes hydrophobic bonding. 2. The second stage leads to visible events like precipitation, agglutination, lysis of cells, killing of Ag, neutralization of toxins, fixation of complement and enhancement of phagocytosis. 3. The third stage includes destruction of Ag or its neutralization.

Salient features of Ag-Ab reactions: 1) Immune complex 2) Specificity of Ag-Ab reaction 3) Binding sites of Ag and Ab

4) Binding forces of Ag and Ab

5) Avidity
6) Bonus effect 7) Cross reaction

Measurement of Ag-Ab reactions: Methods

Many methods are available for the measurement of Ags and Abs participating in the Ag-Ab reactions. Measurement may be done in terms of mass or in terms of titer values. (The degree of dilution of a
substance such as an antibody, reflecting the strength of the solution. )

The two important parameters effecting serological tests are the specificity and the sensitivity .

 A sensitive test should be able to measure even very minute quantities of Ag or Ab. Specificity refers to the ability of the test to detect the reactions between homologous Ags and Abs and not with others.

The Ag-Ab reactions can be detected by the following techniques.

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. Precipitation Agglutination Lysis and complement fixation Neutralization test Flocculation Opsonisation Immunofluorescence Enzyme Immunosorbent assay Radio allegro sorbent test Radio Immunosorbent test Immunoblotting Immunoprecipitation ELISPOT

Precipitation reactions:
When a soluble Ag combines with its Ab in the presence of an electrolyte (NaCl) at a particular temperature and pH, it forms an insoluble precipitate of Ag-Ab complex. The Ab causing precipitation is called Precipitin.

NEUTRALIZATION TESTS Virus neutralization test Toxin-antitoxin ractions FLOCCULATION In these tests Ag-Ab reaction results in visible floccules which do not sediment but remain dispersed in the medium.

AGGLUTINATION REACTIONS:
When a particular Ag is mixed with its Abs in the presence of electrolytes at a suitable temperature and pH, the particles are clumped or agglutinated. The Ab of the serum causes the cellular Ags to form clumps and these are called Agglutinins. The particulate antigens that are aggregated are termed Agglutinogens. This reaction occurs optimally when Ags and Abs react in equivalent proportions.

Mechanism of Agglutination:
1. 2. 3. 4. 5. 6. 7. Slide agglutination Tube agglutination Haemagglutination inhibition test Coombs test (anitglobulin test) Passive agglutination test Brucella agglutination test for Brucellosis Cold agglutinatin test for Pneumonia, Malaria and Trypnosomiasis

ELISA (Enzyme Linked ImmunoSorbent Assay)

ELISA is of 3types.  Indirect ELISA  Sandwich ELISA  Competitive ELISA

ELISPOT