Production and properties of monoclonal antibodies against human IgG isotypes.
Several monoclonal antibodies (MAbs) against human IgG isotypes were obtained by the fusion of myeloma cells with splenocytes from mice immunized with IgG fractions extracted from human plasma. Four MAbs (F7H7, D4F8, B12A8, and E7E10) were selected by an ELISA technique on the basis of their ability to detect one of the four IgG subclasses. Their specificity was checked using a panel of pure myeloma proteins representative of the main allotypes present on IgG isotypes. In addition, two other MAbs (F3E12 and E6D6) were found able to detect specifically kappa or lambda light chains. The immunochemical properties of these MAbs were analyzed mainly in respect to their capacity to detect and to purify the different human IgG isotypes. The following data were obtained: (1) The ability of the MAbs F7H7, D4F8, B12A8, and E7E10 to measure the concentration of each IgG subclass in serum was estimated by an immunocapture ELISA. Results obtained with the new antibodies were compared with several other MAbs recommended by the IUIS/WHO human Immunoglobulins subcommittee. Similar or better results were obtained with the new anti-IgG1, anti-IgG3, and anti-IgG4, MAbs. (2) The same MAbs were tested for their ability to purify a single IgG subclass from IgG preparations and from normal and pathological sera. Fractions containing about 80% of purified IgG1, IgG3, and IgG4 were obtained after one-step immunoaffinity purification. Consequently, these MAbs proved to be useful to detect, to measure and to purify IgG subclasses.
Institut National de la Transfusion Sanguine, Paris, France.
M
M
Hasnaoui
D
D
Blanchard
C
C
Willem
M J
MJ
Loirat
P
P
Lambin
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes