Wood D M, Parent J B, Gazzano-Santoro H, Lim E, Pruyne P T, Watkins J M, Spoor E S, Reardan D T, Trown P W, Conlon P J
Department of Preclinical Science, XOMA Corporation, Berkeley, California.
Circ Shock. 1992 Sep;38(1):55-62.
The murine IgM monoclonal antibody (mAb) E5 was produced by a hybridoma derived from spleen cells of a mouse immunized with the J5 rough mutant of Escherichia coli O111:B4. In a multicenter randomized placebo-controlled clinical trial, E5 has been shown to reduce significantly the mortality and morbidity of patients with Gram-negative sepsis. The characteristics of E5 binding to endotoxin were studied in vitro. We report here the results of binding to an extensive panel of rough lipopolysaccharide (LPS) and lipid A preparations. Using standard immunologic techniques, including enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA), as well as an antibody capture assay using immobilized antibody and a chromogenic Limulus amebocyte lysate (LAL) detection system, E5 was shown to bind to all rough LPS (chemotypes Ra through Re from Salmonella minnesota and E. coli J5) and lipid A preparations tested. E5 displayed a Kd for Ra LPS of approximately 6.5 nM. These results confirm and extend those reported previously and provide evidence that E5 binds specifically to lipid A and to the lipid A moiety of rough LPS.
鼠源IgM单克隆抗体（mAb）E5由一株杂交瘤产生，该杂交瘤源自用大肠杆菌O111:B4的J5粗糙突变株免疫的小鼠脾脏细胞。在一项多中心随机安慰剂对照临床试验中，E5已被证明能显著降低革兰氏阴性菌败血症患者的死亡率和发病率。在体外研究了E5与内毒素结合的特性。我们在此报告了E5与大量粗糙脂多糖（LPS）和脂质A制剂结合的结果。使用包括酶联免疫吸附测定（ELISA）和放射免疫测定（RIA）在内的标准免疫学技术，以及使用固定化抗体和显色鲎试剂检测系统的抗体捕获测定，结果表明E5能与所有测试的粗糙LPS（来自明尼苏达沙门氏菌和大肠杆菌J5的Ra至Re化学型）和脂质A制剂结合。E5对Ra LPS的解离常数（Kd）约为6.5 nM。这些结果证实并扩展了先前报道的结果，并提供了E5特异性结合脂质A和粗糙LPS的脂质A部分的证据。
