Immunological testing Perspective 17.1 Monoclonal

• Monoclonal antibodies • • Quantifying antigen – reactions

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Serology Quantifying antigen – antibody reactions

• Antibodies • Seroconversion or rise in titer • Antibodies detect and identify antigens • Serial dilutions

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Precipitation reactions

• •

Figure 17.2 - Quantitation of immunologic tests

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1 Figure 17.4 Immunodiffusion Figure 17.3 Precipitation reaction

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Agglutination reactions

• Direct • Indirect agglutination •

Figure 17.6 Immunoelectrophoresis

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Direct agglutination Indirect agglutination

• Cross – linking and lattice formation • Soluble antigen is coated onto particles (red bood cells, • Antibodies react with particulate antigens (red blood latex beads) cells, bacteria, fungi) • Allow for visible clumps (agglutination) • Visible clumps • Estimate amount of antibody

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2 tests

• Direct fluorescent antibody test • Indirect fluorescent antibody test

Figure 17.7 - Agglutination reaction

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Figure 17.8 - Direct and indirect fluorescent antibody test Antigen – antibody assays

• (RIA) • Enzyme – linked immunosorbant assay (ELISA) • Western blot

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Enzyme – linked immunosorbant assay Radioimmunoassay (RIA) (ELISA)

• Competitive inhibition assay • Color reaction assay • Measure antigen or antibody • Indirect ELISA • Ex. Measure small amounts of hormones or drugs in a • Direct ELISA clinical sample • Ex. Measure small amounts of IgE antibody (radioallergosorbent test)

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3 Figure 17.9 - Indirect ELISA

Figure 17.1 - Direct ELISA 17-19 17-20

Figure 17.1 Western blot • Measures the binding of complement by an antigen – antibody interaction • Indicator system determine positive or negative reactions

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Figure 17.1 Complement Neutralization test fixation test • Antibody bind to specific antigen (virus, toxin) • Antibody – antigen complex prevents antigen from binding (neutralization)

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4 Cellular test

• Identification of subsets of lymphocytes • Lymphocyte response to mitogens • Cytoxic T – cell function • Cell – mediated immunity to infectious agents

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5