11/26/2014

Antigen Antibody Reactions

Dr Debasis Biswas

Lattice formation

Precipitation Reaction

Lattice: Cross- linking of

Antigen & Antibody molecules

Prozone Zone

Postzone

Zone phenomenon

Precipitation Reaction
Soluble Antigen
Soluble Antibody
Equivalent Proportions
Electrolytes
Temperature
pH

Soluble Antigen
Soluble Antibody
Equivalent Proportions
Electrolytes
Temperature
pH

Antigen- Antibody
Complex Precipitates

Agglutination Reaction

Polyclonal Antibody

Monoclonal Antibody

Antigen Antibody Interaction

Particulate Antigen
Soluble Antibody
Equivalent Proportions
Electrolytes
Temperature
pH

Antigen- Antibody
Complex forms Clumps

Immunodiffusion Reaction
Antigen- Antibody
Complex Precipitates

Radial Immunodiffusion
(Mancini)
Antibody incorporated in gel
Antigen diffuses from well

Flocculation Reaction
Soluble Antigen (low sp gr)
Soluble Antibody
Equivalent Proportions
Electrolytes
Temperature
pH

Antigen- Antibody
Complex Floats

Single Radial Immunodiffusion

(Mancini)
Quantitation of antigen by
measuring diameter of
precipitin ring

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Immunodiffusion Reaction

Immunoelectrophoresis

Double Immunodiffusion
(Ouchterlony)
Antibody & Antigen
diffuse towards each other

Countercurrent Immunoelectrophoresis

Quiz

Rocket
Electrophoresis
Which antigen, H or M, has evoked more antibody formation?
Why?

Agglutination Reaction
Particulate Antigen
Soluble Antibody
Equivalent Proportions
Electrolytes
Temperature
pH

+ +

Antigen- Antibody
Complex forms Clumps

RBC suspension +
S e r i a l D i l u t i o n o f a n t i b o dy

Variants
Latex Agglutination
Hemagglutination

Haemagglutination Reaction

NC: No antibody.
Button formation

Slide Agglutination
Tube Agglutination

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Haemagglutination Inhibition

Latex Agglutination Reaction

Heterophile Agglutination Reaction

Coombs Test

Weil Felix Reaction Rickettsia

Paul Bunnel
Reaction
Cold Agglutinin
Reaction
Streptococcus MG
Agglutination Test

Proteus vulgaris
OX2 OX19
Proteus mirabilis
OXK

Epstein Barr Sheep RBC

virus
Mycoplasma Human RBCs
(Group O)
Mycoplasma Streptococcus MG

Passive Agglutination Test

Incomplete Antibodies:
Bind to particulate antigen
Cannot agglutinate them
Agglutination effected by adding Ab to the incomplete ab

Direct: Infant
Indirect: Mother

ELISA

Target antigen coated on particles

Rose Waaler Test: Hemagglutination Reaction for
Rheumatoid factor

Indirect ELISA: Antibody Detection

Sandwich ELISA: Antigen Detection

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ELISA

Immunofluorescence
Direct IF

Indirect IF
FITC- labelled
Anti- Ig

FITC- labelled Ab
to ag of interest
Incubate antibody
with sample
containing antigen
to be measured

Transfer antigenantibody mixture

to antigen- coated
wells

Ab to ag
of interest

Add enzymeConjugated
Secondary
Antibody

Competitive ELISA: Antigen Detection

Immunofluorescence

Western Blot

Indirect IF
Direct IF

Western Blot

Immunochromatography
Y
: Analyte of interest
: Colloidal gold particles coated with antibody to analyte of interest
: Antibody to analyte of interest
: Antibody to antibody coated on gold particles
T
C
Y

Sample containing analyte of

interest added to test strip

Reaction at T band

Reaction at C band