A simple and accurate microplate assay for the determination of factor XI in plasma.
Traditionally, factor XI has been determined in the clinical laboratory by a modified activated partial thromboplastin time assay (aPTT) with factor XI-deficient plasma as the substrate. Coagulant assays, however, have high coefficients of variation. We previously developed a chromogenic assay for factor XI that required equipment not normally found in a clinical laboratory. We now present a modification of that assay, which is performed in 96-well microplates and can be done in any clinical laboratory or physician's office. Plasma is subjected to a brief acidification to inactivate most of the plasma protease inhibitors. Soybean trypsin inhibitor is included to stabilize the factor XIa that is generated. Kaolin is used as the contact activating surface, and the chromogenic substrate, S-2366, is used to measure the factor XIa that is formed. Results of the assay, performed in three groups of subjects, correlate well with results of the coagulant assay as performed in our laboratory and clearly differentiate between total factor XI deficiency and the deficiency of any of the other contact proteins. Unlike coagulation assays, the chromogenic assay is not influenced by the presence of heparin. Furthermore, it is not affected by lupus anticoagulants, which are antibodies directed against acidic phospholipids. Two plasma samples from patients with acquired factor XI inhibitors showed dissociation between coagulant and amidolytic activity, suggesting that the antibodies were not primarily directed toward the active site of factor XIa, which is responsible for its amidolytic activity. In contrast, patients with severe congenital deficiency of factor XI showed no activity by either assay.
Scott
C F
CF
Department of Medicine, Temple University School of Medicine, Philadelphia, PA 19140.
Colman
R W
RW
D000581:Amidohydrolases / Q000502:physiology; D001777:Blood Coagulation; D003276:Contraceptives, Oral; D005172:Factor XI / Q000032:analysis* / Q000502:physiology; D005173:Factor XI Deficiency / Q000097:blood / Q000235:genetics; D005260:Female; D006405:Hematology / Q000379:methods*; D006493:Heparin / Q000494:pharmacology; D006579:Heterozygote; D006801:Humans; D008297:Male; D010314:Partial Thromboplastin Time; D013997:Time Factors
D003276:Contraceptives, Oral; D006493:Heparin; D005172:Factor XI; D000581:Amidohydrolases