Wolbink G J, Bollen J, Baars J W, ten Berge R J, Swaak A J, Paardekooper J, Hack C E
Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
J Immunol Methods. 1993 Jul 6;163(1):67-76. doi: 10.1016/0022-1759(93)90240-8.
In order to study the activation of the complement system via the classical pathway we have attempted to raise antibodies specific for C4 activation products. Of 20 mouse monoclonal antibodies (mAbs) obtained, one appeared to react with an activation dependent epitope exposed on the activation products C4b, C4bi, C4c (C4b/c) as well as on iC4, but not on native C4. Using this antibody as a capture antibody and polyclonal biotinylated antibodies against C4 as detecting antibodies we developed an ELISA for the quantification of C4b/c in biological fluids. The lower limit of detection was approximately 0.025 nmol C4b/c per litre. Mean C4b/c levels in plasma samples collected from healthy volunteers in tubes containing 10 mM EDTA and 0.05% (w/v) polybrene, final concentrations, appeared to be 30 nmol/l. The potential of the ELISA procedure for evaluating complement activation in clinical samples was demonstrated.
为了研究补体系统通过经典途径的激活，我们试图制备针对C4激活产物的特异性抗体。在获得的20种小鼠单克隆抗体（mAb）中，有一种似乎与C4激活产物C4b、C4bi、C4c（C4b/c）以及iC4上暴露的激活依赖性表位发生反应，但不与天然C4反应。以该抗体作为捕获抗体，以抗C4的多克隆生物素化抗体作为检测抗体，我们开发了一种用于定量生物体液中C4b/c的酶联免疫吸附测定（ELISA）。检测下限约为每升0.025纳摩尔C4b/c。从健康志愿者采集的、含有10 mM乙二胺四乙酸（EDTA）和0.05%（w/v）聚凝胺（最终浓度）的试管中的血浆样本，其C4b/c平均水平似乎为30纳摩尔/升。证明了ELISA程序在评估临床样本中补体激活方面的潜力。
