Misenheimer Tina M, Lasarev Michael R, Kumfer Kraig T, Sheehan John P, Schwartz Bradford S
Morgridge Institute for Research, Madison, Wisconsin, USA.
Department of Biostatistics and Medical Informatics, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, USA.
Res Pract Thromb Haemost. 2024 Feb 5;8(1):102338. doi: 10.1016/j.rpth.2024.102338. eCollection 2024 Jan.
Factor (F)IXa activity has been detected in human plasma and may impact thrombotic risk. Current FIXa activity assays are complex and cumbersome.
To develop a reproducible enzyme-linked immunosorbent assay (ELISA) using a novel monoclonal antibody that detects total FIXa in human plasma.
A monoclonal antibody was raised against the new N-terminus exposed upon activation of FIX to FIXa by cleavage after R226. This antibody is specific for FIXa protease and does not recognize FIX zymogen or FIXα. The antibody was used to develop a FIXa-specific ELISA capable of quantifying total FIXa (free FIXa and FIXa-antithrombin complex) in human plasma. Total FIXa quantified using the ELISA was compared to that of FIXa-antithrombin quantified using modifications of a previously described ELISA.
The FIXa-specific ELISA was reproducible and quantified total FIXa in human plasma. Total FIXa levels correlated with FIXa-antithrombin levels.
A monoclonal antibody was developed that specifically detects human FIXa protease. A FIXa-specific ELISA using the new antibody is capable of reproducibly measuring total FIXa in human plasma (both free FIXa and FIXa-antithrombin). This assay should facilitate the evaluation of total FIXa levels in a variety of clinical circumstances.
已在人血浆中检测到因子（F）IXa活性，其可能影响血栓形成风险。当前的FIXa活性检测方法复杂且繁琐。
使用一种新型单克隆抗体开发一种可重复的酶联免疫吸附测定（ELISA），以检测人血浆中的总FIXa。
制备针对FIX经R226位点裂解激活为FIXa后新暴露的N端的单克隆抗体。该抗体对FIXa蛋白酶具有特异性，不识别FIX酶原或FIXα。使用该抗体开发了一种FIXa特异性ELISA，能够定量人血浆中的总FIXa（游离FIXa和FIXa-抗凝血酶复合物）。将使用ELISA定量的总FIXa与使用先前描述的ELISA的改良方法定量的FIXa-抗凝血酶进行比较。
FIXa特异性ELISA具有可重复性，可定量人血浆中的总FIXa。总FIXa水平与FIXa-抗凝血酶水平相关。
开发了一种特异性检测人FIXa蛋白酶的单克隆抗体。使用该新抗体的FIXa特异性ELISA能够可重复地测量人血浆中的总FIXa（游离FIXa和FIXa-抗凝血酶）。该检测方法应有助于在各种临床情况下评估总FIXa水平。
