Detection of Goodpasture antigen in fractions prepared from collagenase digests of human glomerular basement membrane
Fish, A.J.; Lockwood, M.C.; Wong, M.; Price, R.G.
Clinical and Experimental Immunology 55(1): 58-66
1984
ISSN/ISBN: 0009-9104 PMID: 6319059 Document Number: 229216
Preparations of human glomerular basement membrane (GBM) were digested with collagenase, and a Goodpasture (GP) antigen rich pool from gel filtration column runs was identified by antibody inhibition radioimmunoassay. The components of the GP antigen pool were separated on polyacrylamide gels, and transferred to nitrocellulose sheets by the 'western' blotting technique. The blots were separately reacted with thirteen GP sera as primary antibody, followed by peroxidase labelled goat anti-human IgG and revealed 45-50K (two bands) and 25-28K (one-three bands) components. No corresponding reactivity was observed using convalescent GP sera or other control sera (normal human serum, rapidly progressive glomerulonephritis with or without pulmonary haemorrhage, and lupus erythematosus) as primary antibody.