Limited enzymatic cleavage of pig immunoglobulin and of specific antibodies. V. Monoclonal antibodies to individual fragments and polypeptide chains.
Six hybridomas that produce monoclonal antibodies to different antigenic determinants of heterogeneous pig IgG and of pig anti-Dnp antibodies were obtained by fusion between spleen cells from BALB/c mice immunized with non-specific pig IgG and the myeloma line P3-X63-Ag8.653. Antigenic determinants which correspond to individual monoclonal antibodies were located in the individual domains of the IgG molecule by investigating the interaction of monoclonal antibodies with Fab, Fc, and pFc' fragments and with kappa, lambda, and gamma polypeptide chains. To evaluate the interaction, a quantitative equilibrium competitive radioimmunoassay was developed and [14C]formaldehyde-labelled non-specific pig IgG served as labelled antigen. Antibodies PGG-01, PGG-04, and PGG-06 were shown to be directed against determinants in the C lambda domain, antibody PGG-03 very probably against the determinant in the C chi domain, and antibodies PGG-02 and PGG-05 against determinants in the CH3 domain. Antibodies PGG-02 and PGG-05 are highly selective for IgG subpopulations; when combined, they interact with no more than 42% of the heterogeneous IgG population. These two antibodies can be used as a basis for preparing a complete set of reagents for identification of individual subclasses.
D019297:2,4-Dinitrophenol; D000818:Animals; D000911:Antibodies, Monoclonal; D004140:Dinitrophenols; D000939:Epitopes / Q000032:analysis; D005260:Female; D007074:Immunoglobulin G / Q000032:analysis* / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010434:Pepsin A / Q000378:metabolism; D010446:Peptide Fragments / Q000276:immunology*; D013552:Swine; D014357:Trypsin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D004140:Dinitrophenols; D000939:Epitopes; D007074:Immunoglobulin G; D010446:Peptide Fragments; D014357:Trypsin; D010434:Pepsin A; D019297:2,4-Dinitrophenol