Precipitation and Agglutination Reactions Flashcards
Antigen–Antibody Binding: Affinity
Is the initial attraction force between a Fab site on an antibody molecule and an epitope or determinant site on an antigen
strength of attraction depends on the specificity of the antibody for a particular antigen
Antigen–Antibody Binding: Avidity
the sum of the attractive forces between an antigen and an antibody that keeps the molecules together
Is a measure of the overall stability of an antigen–antibody complex
Precipitation Reactions
Involve combining soluble antigen with soluble antibody to produce insoluble complexes that are visible
What is required for antigen and antibody in Precipitation reactions?
Multiple binding sites for one another
Equal relative concentration of each
Zone of Equivalence
Number of multi-valent sites of antigen and antibody are approximately equal
Prozone
antibody excess
Antigen combines with only one or two antibody molecules
False-negative reactions may occur as a result of high antibody concentration
Postzone
antigen excess
The presence of a small amount of antibody may be obscured, causing false-negative results
Nephelometry
Light that is scattered at an angle is measured, indicating the amount of antigen or antibody present
Radial immunodiffusion
Antigen diffuses out into gel that is infused with antibody. Measurement of the radius indicates concentration of antigen
Ouchterlony double diffusion
Both antigen and antibody diffuse out from wells in a gel. The lines of precipitate formed indicate the relationship of antigens
Immuno-electrophoresis
Electrophoresis of serum is followed by diffusion of antibody from the wells
Immunofixation electrophoresis
Electrophoresis of serum is followed by direct application of antibody to the gel
Difference b/n Ouchterlony and Radial immunodiffusion
Radial immunodiffusion: single diffusion
Ouchterlony: Double diffusion
Ouchterlony result interpretation
Arch: Identity
X shape: non identity
A line with a spur: Partial Identity
Agglutination
Is the visible aggregation of particles resulting from combination with specific antibody