antigen-antibody interactions. serology - in vitro demonstration of ag/ab reaction ag/ab reaction =...
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Antigen-Antibody Interactions
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Serology - in vitro demonstration of Ag/Ab reaction
Ag/Ab reaction = reversible1. Primary interactions
1) Electrostatic interactions2) Hydrogen bond3) Hydrophobic interactions4) van de Waals force
2. Secondary interactionsPrecipitationAgglutinationComplement activation reactions
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Association constant
Ab + H AbH (binding of hapten)
K = [AbH]/[Ab][H] (association constant between reactants)
Affinity = binding of Ab with epitope characterized by association constant
Avidity = binding energy between Ab & multivalent Ag IgM > IgG
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Figure 5.1Reactions between antibody or antibody fragments and antigens or hapten.
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Figure 5.2
A representation of the anti-immunoglobulin (Coombs) test.
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Agglutination inhibition (Early home pregnancy test kit)
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Figure 5.3
A representation of the precipitin reaction.
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Figure 5.4Gel diffusion by antibodies and a single antigen (A) and antibodies to
antigens 1, 2, 3, and their respective antigens (B).
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Figure 5.5Double gel-diffusion patterns showing pattern of identity (left), pattern of nonidentity (center), and pattern of partial identity (right).
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Figure 5.6Radial diffusion, A, B, C, D, and E represent known concentrations of antigen; F and G represent unknown concentrations that can be
determined from the graph.
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Figure 5.7
Patterns of immunoelectrophoresis of serum proteins.
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Figure 5.8Western blots of serum samples from two HIV-infected individuals and one control subject.
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Figure 5.9Amount of label bound to antibody after incubation of constant amounts of antibody and labeled antigen.
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Figure 5.10Radioimmunoassay, based on the competition of nonlabeled and labeled antigens for antibody.
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Solid phase radioimmunoassay (RIA) todetect hepatitis B virus in blood serum
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Figure 5.11A representative ELISA using a well coated directly with antigen
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Immunoprecipitation
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Immunofluorescence
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Figure 5.12A schematic representation of a fluorescence-
activated cell sorter (FACS).
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Flow cytometry & leukemia typing
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Figure 5.13
A schematic representation of the production of monoclonal antibodies.
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Figure 5.14A general procedure for producing transgenic mice.