Antigen-antibody reaction

 Antigens and antibodies combine with each other

specifically and in an observable manner.
 In the body, they form the basis of antibody mediated
immunity in infectious diseases, or hypersensitivity and
autoimmune diseases.
 In laboratory, they help in diagnosis of infections, in
epidemiological surveys, in the identification of infectious
agents, enzymes.
Antigen – antibody reactions in vitro are known as
serological reactions.
Stages of Ag – Ab reactions
Primary stage:
Initial interaction between Ag & Ab – invisible
Rapid, occurs at low temperatures & obeys the
general laws of physical chemistry &
thermodynamics.
Reaction is reversible.
Ag & Ab is bound to each other by weak Van der
Waal’s forces, Ionic bonds & Hydrogen bonding.
Secondary stage:
Demonstrable events – Precipitation, agglutination,
lysis of cells, killing of live antigens, neutralization of
toxins, complement fixation, immobilization of motile
organisms & enhancement of phagocytosis.

Precipitin – Ab participate in precipitation

Agglutinin - Ab participate in agglutination
Precipitinogen – Ag participate in precipitation
Agglutinogen - Ag participate in agglutination
Tertiary stage:

Includes neutralization or destruction of injurious agents

or tissue damage.
Also includes humoral immunity against infectious
diseases as well as clinical allergy & other immunological
diseases.
 GENERAL FEATURES OF Ag – Ab REACTIONS

1. The reaction is specific.

2. Entire molecules react and not the fragments
3. There is no denaturation of the antigen or antibody
during the reaction.
4. The combination occurs at the surface. So surface
antigens are immunologically relevant.
5. The combination is firm but reversible. The firmness
is influenced by the affinity & avidity of the
reaction.
Affinity – refers to the intensity of attraction between the
antigen & antibody molecules. It is the function of
closeness of fit between the epitope & antigen binding
region of its Ab.
Avidity – strength of the bond after the formation Ag-Ab
complexes.

6. Both Ags & Abs participate in the formation of

agglutinates or precipitates.
7. Antigens & antibodies can combine in varying
proportions. Both Ags & Abs are multivalent.
MEASUREMENT OF ANTIGEN & ANTIBODY

Measurement may be in terms of mass or more commonly

as units or titre.
The Antibody titre of a serum is the highest dilution of the
serum which shows an observable reactions with the
antigen in a particular test.
Two important parameters in serological tests – Sensitivity &
Specificity
Sensitivity: The ability of the test to detect even very minute
quantities of antigen or antibody.
When the test is highly sensitive, false negative results
may be absent or minimal.
Specificity: The ability of the test to detect reactions between
homologous Ags & Abs only, and with no other.
In highly specific test, false positive reactions are absent
or minimal.
 PRECIPITATION REACTION

PRINCIPLE: When a soluble Ag combines with its Ab in

the presence of electrolytes (NaCl) at a suitable
temperature & pH, the Ag-Ab complex forms an
insoluble precipitate.
When instead of sedimenting, the precipitate remains
suspended as floccules – Flocculation reaction
Precipitation can take place in liquid media or in gels
such as agar, agarose or polyacrylamide.
Antigen= immunogen: any foreign substance which, when
introduced will evoke a specific immune response.
In terms of infectious diseases, the following may act as
antigens:
 Microbial structures (cell walls, capsules, flagella, pili,
viral capsids, envelope-associated glycoproteins, etc.).
 Microbial exotoxins.
Certain noninfectious materials may also act as
antigens if they are recognized as "nonself" by the
body. These include:
1. Allergens.
2. Foreign tissues and cells.
3. The body's own cells that the body fails to recognize
as "normal self" as cancer cells.
Antibodies or immunoglobulins are
specific protein configurations
produced by B-lymphocytes and
plasma cells in response to a specific
antigen and capable of reacting with
that antigen.

Antibody structure (IgG)

Antigen-antibody reactions can be used in the
laboratory to identify unknown
microorganisms
 Antigen antibodies reaction

1. agglutination
2. precipitation
3. complement fixation
4. Toxin antitoxin neutralization
5. virus neutralization
6. Immunefluorescence
7. Flow cytometry
8. Enzyme linked immunesorbant assay (ELISA)
9. Radioimmunoassay
Serologic testing may be used in the clinical laboratory in
two distinct ways:

To identify unknown antigens (such as microorganisms).

To detect antibodies being made against a specific antigen

in the patient's serum.

 Agglutination
Antigen: suspension of microorganisms,

cells or latex particles coated with antigens.

Antibodies: specific anti serum

Reaction …………………… Agglutination

or clumping.

If one (antigen or antibody) is known, the

reaction can be used for identification of the
other.
Factors affect antigens antibodies reaction
ICT
Thanks