Precipitation Reactions in Fluids Yield a Precipitin Curve

A quantitative precipitation reaction can be performed by placing a constant amount of antibody in a series of tubes and adding increasing amounts of antigen to the tubes. At one time this method was used to measure the amount of antigen or antibody present in a sample of interest. After the precipitate forms, each tube is centrifuged to pellet the precipitate, the supernatant is poured off, and the amount of precipitate is measured. Plotting the amount of precipitate against increasing antigen concentrations yields a precipitin curve. As Figure 6-4b shows, excess of either antibody or antigen interferes with maximal precipitation, which occurs in the so-called equivalence zone, within which the ratio of antibody to antigen is optimal. As a large multimolecular lattice is formed at equivalence, the complex increases in size and precipitates out of solution. As shown in Figure 6-4, under conditions of antibody excess or antigen excess, extensive lattices do not form and precipitation is inhibited. Although the quantitative precipitation reaction is seldom used exper imentally today, the principles of antigen excess, antibody excess, and equivalence apply to many Ag-Ab reactions.

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