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An immunoradiometric assay is described for the determination of human anti-idiotypic anti-B72.3 IgG. The latter is formed in ovarian cancer patients after treatment with the murine monoclonal antibody B72.3, which is directed against the tumour-associated glycoprotein 72 (TAG-72). A gel coupled with Fc-specific anti-human IgG antibodies is used as a solid phase for the extraction of serum IgG. The anti-B72.3 IgG is then specifically detected by incubation with radiolabelled B72.3 detector antibodies. Calibration standards were prepared from serum obtained from a patient repeatedly treated with B72.3 antibodies. The concentration of anti-idiotypic anti-B72.3 antibodies was expressed as TAG-72-like arb.units/1. The assay performed with two 60-minute incubation steps is characterized by a high sensitivity (detection limit: 3 x 10(3) arb.units/1) and precision (coefficients of variation: intra-assay = 6.4% and 5.8% at 80 x 10(3) arb. units/1 and 217 x 10(3) arb.units/1, inter-assay = 8.7% and 7.1% at 91 x 10(3) arb.units/1 and 212 x 10(3) arb.units/1) and a good linearity of dilution (recovery after dilution between 99% and 107%). The assay is more specific than previously described methods; no interference was observed by TAG-72 up to 3.3 x 10(7) arb.units/1. Also, non-specific human anti-mouse antibodies did not cross-react up to 34.8 mg/l. The test may be modified for detection of anti-idiotypic antibodies, which are formed after treatment with other monoclonal antibodies.

clinical chemistry and laboratory medicine

A Simple and Sensitive Assay for Determination of Human Anti-Idiotypic Anti-B72.3 Antibodies, which Is not Affected by the Presence of Tumour-Associated Glycoprotein 72