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Primary Biliary Cirrhosis (PBC): Characterization of a Monoclonal Antibody (PBC‐MoAb) having Specificity Identical with Disease‐Associated Auto‐antibodies
Author(s) -
BJÖRKLAND A.,
MENDELHARTVIG I.,
NELSON B. D.,
TÖTTERMAN T. H.
Publication year - 1991
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1991.tb02549.x
Subject(s) - epitope , primary biliary cirrhosis , monoclonal antibody , autoantibody , antibody , western blot , pyruvate dehydrogenase complex , microbiology and biotechnology , epitope mapping , blot , linear epitope , chemistry , enzyme , biology , biochemistry , immunology , gene
We have raised a monoclonal antibody (PBC‐MoAb) directed against mitochondria which resembles patent anti‐mitochondrial autoantibodies (AMA) (M2 type) in several respects. The reaction pattern of PBC‐MoAb was characterized by western blot experiments, immunoaffinity purification and enzyme inhibition studies. PBC‐MoAb reacts specifically with an epitope on the E2 suburil of pyruvate dehydrogenase (dihydrolipoamide acyltransferase) which is essential for enzymatic activity. This was shown as follows: (I) PBC‐MoAb. like PBC‐AMA, completely inhibited PDH enzyme activity and reacted weakly with OGDII; (2) PBC‐MoAb bound strongly to the E2 subunit in western blots, with weaker binding to a doublet of about 56 kDa: and O) in immunosorbent experiments, PBC‐MoAb absorbed most (>95%) 0of the AMA reactive material found in solubilized mitochondria. The present data together with earlier findings that the majority of PBC patient autoantibodies bind to epitopes defined by the PBC‐MoAb, makes this antibody a valuable tool for characterizing the major PBC‐associated epitope on PDH‐E2 and localizing this epitope in liver tissue.