Solid-phase radioimmunoassay of immunoglobulins G, A and M: applicability in analysis of sucrose gradients.
A simple and sensitive solid-phase radioimmunoassay for the detection of immunoglobulins G, A and M in sucrose gradients is described. The solid-phase consisted of immunoglobulins adsorbed to polystyrene tubes. Using buffers without detergent and 125I-labeled sheep anti-rabbit IgG as radioligand, the assay was able to detect 0.8 ng per tube in the IgG assay and 1.6 ng per tube in the IgA and IgM assays. Standard curves with antigen dissolved in 10% and 32% sucrose were superimposable and did not deviate from standard curves with antigen dissolved in buffer without sucrose. Using these techniques on ultracentrifugation samples from patients with systemic lupus erythematosus, Schönlein-Henoch nephritis and IgA glorulonephritis it was possible to detect both immunoglobulin fragments and immunoglobulin aggregates at the same time without prior dialysis of the samples.
D000818:Animals; D002499:Centrifugation, Density Gradient; D006801:Humans; D007105:Immune Complex Diseases / Q000097:blood; D007070:Immunoglobulin A / Q000032:analysis*; D007074:Immunoglobulin G / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis*; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D014461:Ultracentrifugation / Q000379:methods
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M