Lecture 3

Antibody-Antigen Reactions

Binding of the epitope in

the antigen binding site
GOOD FIT

POOR FIT

antibody combining site

antigen determinant
high attraction
low repulsion

high repulsion
low attraction

AntibodyAvidity

Multiple interactions between antigen binding

sites and epitopes
Greater than additive
Similar to Velcro
More relevant to biologic systems than affinity

Effect of multivalent
interactions
antibody

Fab

IgG

IgG

IgM

effective antibody valence

up to 10

antigen valence

equilibrium constant (L/M)

10 4

10 4

10 7

10 11

advantage of multi-valence

10 3 -fold

10 7 -fold

affinity

affinity

avidity

avidity

definition of bindng

intrinsic affinity

functional affinity

Biological Consequences of
Antibody Affinity/Avidity

Neutralization of toxins
Complement activation
Immune elimination of antigen
Virus neutralization
More intense immune complex disease in animals
higher levels of circulating antigen-antibody complexes
more intense localization of immune complexes on
basement membranes.
more severe impairment of organ function

Cross-Reaction
A

B C

Immunogen

Immunize

anti-A
A

B C

anti-B

anti-C

anti-D

Reactivity with
rabbit
antiserum

Cross-reactive antigen

Antigen-Antibody
Interactions
Quality and quantity are important
in resolution of disease
May contribute to pathology
Useful in immunological assays

Immune Precipitation

Antigen
Antibody

Agglutination
RBC

IgM Antibody

RBC

RBC

RBC

RBC

RBC

RBC

RBC

IgG Antibody
RBC

RBC

RBC

Enzyme (ELISA) Immunoassay

Enzyme conjugated
to anti-Ig antibody
(second antibody)

Add
substrate
for enzyme

anti-antigen
antibody

Antigen

Complement activation

A system of plasma proteins that interact with

Antigen/antibody complexes
Pathogen surface motifs (alternative and lectin pathways

Activation of complement results in

Chemo-attraction of inflammatory cells

Peptide mediators of inflammation (anaphylatoxins)

Increased blood vessel permeability

Smooth muscle contraction
Mast cell degranulation

Opsonization of pathogens (enhances phagocytosis)

Killing of pathogens (membrane attack complex)

Overview of the Complement

Cascade

Effector
Actions of
Complement

Terms to remember

Affinity
Agglutination
Avidity
Complement

Classical pathway
Alternative (properdin)
pathway

Cross-reaction
Epitope/Antigenic
Determinant
Precipitation
Second antibody
Specificity
Valence