Production of Monoclonal Antibodies Against IgG Allotypes and Their Use in Dot Immunobinding
IgG heavy chain allotypes, GM, are extremely useful in population genetics and in cases of disputed paternity. Unfortunately, the longstanding shortage of typing reagents hampers the widespread use of GM allotyping. Moreover, the cumbersome hemagglutination inhibition (HAI) method of typing is still generally used: 0, D(+) red cells coated with GM-positive anti-D antibodies are agglutinated by the anti-GM serum; prior addition of a serum sample to the anti-GM serum inhibits the hemagglutination if the test serum contains that particular allotype. Despite its ingenuity, HAI poses several problems: (1) The shelf life of coated red cells is relatively short; thus the cumbersome step of coating must be repeated at frequent intervals. (2) Since human anti-GM sera are scarce and of low titer, a large number of healty individuals need continually to be screened for anti-GM antibodies so that the stock of antisera may not be exhausted. The screening is laborious and presupposes the availability of GM allotyping control sera and anti-D antibodies of various GM specificities which are extremely difficult to obtain from certain racial groups. (3) The occaisional occurrence in serum samples of antibodies to IgG and/or red cell antigens invalidates the HAI test unless they are absorbed by a time-consuming procedure. (4) The HAI test can not be used for the quantitative analysis of GM allotypes. All of these problems are solved by the use of monoclonal antibodies (MAbs) in enzyme-linked immunosorbent assay (ELISA) or dot immunobinding (DIB) (Fletcher et al. 1983; Kishida and Tamaki 1984; Bird et al. 1984; Francois-Gerard and Hoste 1987; Kishida et al. 1988; Weston-Kirkegaard et al. 1988; de Lange et al. 1989).
KeywordsCapture ELISA Serial Double Dilution Popliteal Lymph Node Cell Dispute Paternity Mouse IgG1 MAbs
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