Human IgG subclass assays using a novel assay method.
To facilitate assays for human IgG subclasses, we have adapted a novel assay method (particle concentration fluorescence immunoassay) (Jolley et al., 1984; MacCrindle et al., 1985) using one polyclonal and several monoclonal antibodies for human IgG subclasses. The advantages of this new assay over previously described methods are sensitivity (0.3-3 micrograms/ml), and automated measurement of multiple samples in a short time (2 h). We found that a monkey antibody for IgG2 and monoclonal antibodies for IgG1, IgG4b epitope, and IgG3 can be adapted to this method. We evaluated 7 different antibodies to IgG4 without finding a suitable monoclonal antibody for this assay method. Several of these IgG4 hybridoma antibodies, however, could be used in a competitive radioimmunoassay using polyvinyl microtiter plates. The usefulness of a monoclonal antibody to the IgG4b epitope was evaluated because no suitable monoclonal antibodies for IgG2 are available. Because IgG4 levels are usually much smaller than IgG2 levels, and the IgG4b epitope is expressed on all IgG2 alleles and only some IgG4 alleles (Kunkel et al., 1970), the antibody for IgG4b is potentially useful to screen a large number of samples for IgG2 deficiency. However, when the monoclonal antibody for IgG4b was compared with an IgG2 specific antibody produced in a monkey, the IgG4b antibody could identify only about half of the patients with known IgG2 deficiency.
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D005455:Fluorescent Antibody Technique; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000145:classification*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G