Changes of immunoreactivity in alpha 1-antitrypsin in patients with autoimmune diseases.
Recent studies from this laboratory have shown that a monoclonal antibody prepared against a specific epitope on alpha 1-antitrypsin is a valuable diagnostic marker for autoimmune conditions. In the present study we have further characterized this monoclonal antibody and reassessed its diagnostic value in screening samples from patients with various autoimmune conditions. alpha 1-Antitrypsin was micropurified from patients with selected autoimmune conditions and from normal donors. The purified alpha 1-antitrypsin isolated from patients with autoimmune conditions and normal donors was deglycosylated using both a mixture of exoglycosidases and endoglycosidase F. The immunoreactivity of the native and deglycosylated alpha 1-antitrypsin was examined using both a monoclonal antibody and a polyclonal antibody in enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA), respectively. It was noted that alpha 1-antitrypsin isolated from patients with autoimmune diseases generated a displacement curve dissimilar to alpha 1-antitrypsin purified from normal donors or alpha 1-antitrypsin from patients with autoimmune diseases subjected to deglycosylation when these samples were examined by ELISA using the monoclonal antibody. However, when the polyclonal antibody was used for these studies, no difference was found between the native and deglycosylated alpha 1-antitrypsin suggesting that the monoclonal antibody recognized an epitope not detectable by the polyclonal antibody.(ABSTRACT TRUNCATED AT 250 WORDS)
Saso
L
L
Population Council, New York, New York 10021.
Silvestrini
B
B
Lahita
R
R
Cheng
C Y
CY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001327:Autoimmune Diseases / Q000097:blood / Q000175:diagnosis / Q000276:immunology*; D015415:Biomarkers; D002272:Carcinoembryonic Antigen / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D015151:Immunoblotting; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008403:Mass Screening; D011489:Protein Denaturation; D011863:Radioimmunoassay; D051381:Rats; D012859:Sjogren's Syndrome / Q000276:immunology; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000187:drug effects / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D002272:Carcinoembryonic Antigen; D000939:Epitopes; D000515:alpha 1-Antitrypsin; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase