Method for testing antiserum titer and avidity in nephelometric systems.
Antiserum performance in a nephelometric system can be characterized by parameters derived from measuring reaction rates. The characterization process is derived from a series of dose-response curves (elicited nephelometric response vs antigen concentration) generated from various dilutions of the antiserum being tested. Antiserum titer can then be calculated by plotting the antigen concentration found at one-half the maximum nephelometric response (Hmax) of each dose-response curve (C50) vs the corresponding antiserum dilution. Antiserum avidity can be calculated by plotting Hmax against its corresponding antiserum concentration. After general expressions are determined for C50 and Hmax vs antiserum concentration, a single dose-response curve suffices for characterizing antisera with respect to titer and avidity. Direct evidence is provided for the validity of C50 and Hmax as measures of titer and avidity by correlating these parameters with antiserum binding strength and with the number of antibodies eluted from immobilized antigen. This method can be applied to evaluate and compare different antiserum lots having the same specificity, to identify reagent inadequacies by comparing antisera of different specificity, and to predict the optimal antiserum dilution to use in performing an assay.
D000936:Antigen-Antibody Complex; D000941:Antigens / Q000032:analysis*; D006801:Humans; D007106:Immune Sera / Q000032:analysis*; D007074:Immunoglobulin G / Q000032:analysis; D007700:Kinetics; D009391:Nephelometry and Turbidimetry / Q000379:methods; D009961:Orosomucoid / Q000032:analysis; D000515:alpha 1-Antitrypsin / Q000032:analysis
D000936:Antigen-Antibody Complex; D000941:Antigens; D007106:Immune Sera; D007074:Immunoglobulin G; D009961:Orosomucoid; D000515:alpha 1-Antitrypsin