Light scattering by immune complexes

When the precipitin reaction is performed under proper conditions, the amount of light scattered will be directly related to the reactant concentration. A prerequisite for this is that the reaction is performed with one of the reactants, usually the antibody, in excess. A given antigen concentration will then result in a given amount of immune complexes and consequently a given amount of scattered light. At high antigen concentrations, however, when the precipitin curve approaches the equivalence zone or eventually proceeds into the antigen excess zone, the scatter response is no longer linear.

The light-scattering properties vary in different patients' sera, and in order to circumvent this problem, subtraction of individual serum blank values is required for each separate analysis. High blank values may be due to lipemic sera, or to the existence of endogenous immune complexes, for example. Aggregated immunoglobulins, particularly after repeated freezing and thawing, may also contribute to an increased background value. The problem of varying blank values is one reason for the greater sensitivity of protein quantitation in other body fluids such as urine, for example, as compared to serum. From the practical point of view it is usually the problem of high blank values for serum samples which restricts the sensitivity in nephelometric analyses.

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