Monoclonal antibodies Serology Quantifying antigen ...faculty.mercer.edu/green_gr/Biosciences...
Transcript of Monoclonal antibodies Serology Quantifying antigen ...faculty.mercer.edu/green_gr/Biosciences...
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Immunological testing
• Monoclonal antibodies• Serology• Quantifying antigen – antibody reactions
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Perspective 17.1Monoclonal Antibodies
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Serology
• Antibodies• Antibodies detect and identify antigens
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Quantifying antigen – antibody reactions
• Seroconversion or rise in titer• Serial dilutions
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Figure 17.2 - Quantitation of immunologic tests
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Precipitation reactions
• Immunodiffusion• Immunoelectrophoresis
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Figure 17.3Precipitation reaction
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Figure 17.4Immunodiffusion
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Figure 17.6Immunoelectrophoresis
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Agglutination reactions
• Direct agglutination• Indirect agglutination• Hemagglutination
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Direct agglutination
• Cross – linking and lattice formation• Antibodies react with particulate antigens (red blood
cells, bacteria, fungi)• Visible clumps • Estimate amount of antibody
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Indirect agglutination
• Soluble antigen is coated onto particles (red bood cells, latex beads)
• Allow for visible clumps (agglutination)
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Figure 17.7 - Agglutination reaction
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Immunofluorescence tests
• Direct fluorescent antibody test• Indirect fluorescent antibody test
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Figure 17.8 - Direct andindirect fluorescent antibody test
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Antigen – antibody assays
• Radioimmunoassay (RIA)• Enzyme – linked immunosorbant assay (ELISA)• Western blot
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Radioimmunoassay (RIA)
• Competitive inhibition assay• Measure antigen or antibody• Ex. Measure small amounts of hormones or drugs in a
clinical sample• Ex. Measure small amounts of IgE antibody
(radioallergosorbent test)
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Enzyme – linked immunosorbant assay (ELISA)
• Color reaction assay• Indirect ELISA• Direct ELISA
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Figure 17.9 - Indirect ELISA
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Figure 17.1Western blot
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Complement fixation test
• Measures the binding of complement by an antigen –antibody interaction
• Indicator system determine positive or negative reactions
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Figure 17.1Complementfixation test
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Neutralization test
• Antibody bind to specific antigen (virus, toxin)• Antibody – antigen complex prevents antigen from
binding (neutralization)
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Cellular immunology test
• Identification of subsets of lymphocytes• Lymphocyte response to mitogens• Cytoxic T – cell function• Cell – mediated immunity to infectious agents