The detection of circulating antibodies to human glomerular basement membrane

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Humoral antibodies specific for glomerular basement membrane (GBM) of kidney are of major importance in the immunopathogenesis of Goodpasture's syndrome and some forms of human and experimental glomerulonephritis. Circulating anti-GBM antibodies have been detected by radioimmunochemical methods in the sera of patients with Goodpasture's syndrome and linear deposit glomerulonephritis. Human glomerular basement membrane was isolated, purified, and an antigenic component extracted with 8M urea. The 8M urea extract was radiolabelled, separated by gel filtration chromatography, and employed as GBM antigen for detection of binding in a double antibody radioimmunoassay. A series of experiments describe conditions and handling procedures in a radioimmunoassay that yield high binding of the radiolabelled preparation of GBM to sera from patients while binding to normal sera remained low. [...]