An automated microassay for enzyme inhibitory effects of M2 antibodies in primary biliary cirrhosis.
In primary biliary cirrhosis (PBC), autoantibodies are produced to the M2 group of mitochondrial antigens, of which a major constituent is the E2 subunit of the pyruvate dehydrogenase complex (PDC). These antibodies, in addition to conventional reactivities with PDC, characteristically inhibit the catalytic function of PDC in vitro, as judged by a macroinhibition assay based on spectrophotometry. We describe a microinhibition assay adapted for microtitre plates and for an automated readout of results by an ELISA plate reader. We show that this microassay has a sensitivity similar to that of the macroassay. In a study of 83 sera, from PBC and other diseases, the inhibitory microassay proved specific for PBC. This automated inhibitory microassay for PBC-sera could become a primary laboratory procedure for the diagnosis of PBC, particularly because it may identify antibodies to the actual autoepitope on the PDC-E2.
Teoh
K L
KL
Centre for Molecular Biology and Medicine, Monash University, Clayton, Australia.
Rowley
M J
MJ
Mackay
I R
IR
D000818:Animals; D001323:Autoantibodies / Q000097:blood* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006801:Humans; D008105:Liver Cirrhosis, Biliary / Q000175:diagnosis / Q000276:immunology*; D008930:Mitochondria, Liver / Q000201:enzymology / Q000276:immunology*; D011768:Pyruvate Dehydrogenase Complex / Q000037:antagonists & inhibitors* / Q000276:immunology; D013053:Spectrophotometry
D001323:Autoantibodies; D000939:Epitopes; D011768:Pyruvate Dehydrogenase Complex