Katayama M, Hirai S, Kato I, Titani K
Cell Technology Reagent Section, Biotechnology Research Laboratories, Takara Shuzo Co., Ltd., Shiga, Japan.
Clin Biochem. 1994 Apr;27(2):123-31. doi: 10.1016/0009-9120(94)90022-1.
High-molecular-weight von Willebrand factor (vWf) multimers were separated from their smaller multimers by molecular-sieve chromatography and were measured by several sandwich enzyme-linked immunosorbent assays (ELISA) with monoclonal antibodies (moABs) against human vWf. The epitopes of these moABs were mapped partially using fragments generated by V-8 protease digests of native vWf. Large multimers were more immunoreactive with the sandwich ELISA using immobilized VW28-1 and enzyme-labeled VW92-3 than with any other ELISA. The epitopes recognized by these two moABs were sensitive to trypsin and plasmin digestion, and the other moABs appeared to be reactive with the extensively digested antigen. Relative reactivities of this ELISA to plasma vWf multimers in diabetes mellitus were significantly reduced compared to those in healthy subjects. These data demonstrate that molecular abnormalities of vWf circulating in diabetic patients may be caused by some plasma proteases that increase in the microcirculation of patients with diabetic vascular diseases.
通过分子筛色谱法将高分子量血管性血友病因子（vWf）多聚体与其较小的多聚体分离，并用几种针对人vWf的单克隆抗体（moABs）的夹心酶联免疫吸附测定（ELISA）进行检测。使用天然vWf的V-8蛋白酶消化产生的片段对这些moABs的表位进行了部分定位。与任何其他ELISA相比，使用固定化VW28-1和酶标记的VW92-3的夹心ELISA对大多聚体的免疫反应性更高。这两种moABs识别的表位对胰蛋白酶和纤溶酶消化敏感，而其他moABs似乎与广泛消化的抗原发生反应。与健康受试者相比，该ELISA对糖尿病患者血浆vWf多聚体的相对反应性显著降低。这些数据表明，糖尿病患者循环中vWf的分子异常可能是由糖尿病血管疾病患者微循环中增加的一些血浆蛋白酶引起的。
