A solid-phase antibody capture assay for the measurement of C1-inhibitor consumption in vivo.
C1-inhibitor (C1-Inh) is a serine esterase proteinase inhibitor (serpin) which plays an important role in regulating serine proteinases of the early inflammatory response. In this study, we describe a novel and versatile polyclonal antibody capture assay to examine C1-Inh consumption in vivo. This assay has advantages over previously described methods of measuring C1-Inh consumption as it allows the assessment of the relative amounts of native, complexed and cleaved inhibitor circulating in plasma. By using polyclonal antibodies specific for other complement proteins, the C1-Inh capture assay was adapted to measure in vivo activation of C3, C4 and factor B. C1-Inh consumption and complement activation were examined in the plasma of 21 normal individuals, 24 individuals with systemic lupus erythematosus (SLE), nine individuals with adult respiratory distress syndrome (ARDS) and in the paired plasma and synovial fluid from 18 patients with rheumatoid arthritis (RA). The C1-Inh capture assay revealed native, cleaved and complexed C1-Inh migrating at 115 kDa, 96 kDa and 209-225 kDa respectively, in normal plasma. C1-Inh consumption was increased in the plasma of all the inflammatory disorders examined, in comparison to normal plasma. It is proposed that this serpin capture assay could be adapted to the study of serpin involvement in a wide variety of inflammatory disorders.
Wallace
E M
EM
Department of Immunology, Trinity College Medical School, St James's Hospital, Dublin, Ireland.
Feighery
C
C
Jackson
J
J
D001172:Arthritis, Rheumatoid / Q000378:metabolism; D003167:Complement Activation; D003174:Complement C1 Inactivator Proteins / Q000032:analysis / Q000378:metabolism*; D003176:Complement C3 / Q000032:analysis; D003181:Complement C4 / Q000032:analysis; D011415:Complement Factor B / Q000032:analysis; D003165:Complement System Proteins / Q000032:analysis; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008180:Lupus Erythematosus, Systemic / Q000097:blood; D012128:Respiratory Distress Syndrome / Q000097:blood; D013582:Synovial Fluid / Q000378:metabolism
D003174:Complement C1 Inactivator Proteins; D003176:Complement C3; D003181:Complement C4; D003165:Complement System Proteins; D011415:Complement Factor B