Department of Cancer and Inflammation, Institute of Molecular Medicine, Faculty of Health Science, University of Southern Denmark, Odense, Denmark.
J Immunol Methods. 2010 Oct 31;362(1-2):142-50. doi: 10.1016/j.jim.2010.09.024. Epub 2010 Sep 24.
There is a general need for markers of systemic inflammation in acute or chronic diseases, where complement activation is involved. Available methods to monitor complement activation are elaborate and of low sensitivity; they include haemolytic assays (CH50), quantification of fluid phase terminal complex (C5b-C9) and quantification of complement split products by precipitation-in-gel techniques (e.g. C3d). We have developed a mouse monoclonal antibody (mAb) that is able to detect fluid phase C3c without interference from other products generated from the complement component C3. The C3c specific mAb was tested in different ELISA combinations with various types of in vitro activated sera and with plasma or serum samples from factor I deficient patients. The specificity of the mAb was evaluated in immunoprecipitation techniques and by analysis of eluted fragments of C3 after immunoaffinity chromatography. The C3c mAb was confirmed to be C3c specific, as it showed no cross-reactivity with native (un-cleaved) C3, with C3b, iC3b, or with C3d. Also, no significant reaction was observed with C3 fragments in factor I deficient sera or plasma. This antibody forms the basis for the generation of a robust ELISA that allows for a quick and reliable evaluation of complement activation and consumption as a marker for inflammatory processes. We established the C3c plasma range in 100 healthy Danish blood donors with a mean of 3.47 μg/ml and a range of 2.12-4.92 μg/ml. We believe that such an antibody might be of potential value in the assessment of in vivo complement activity during the inflammatory processes.
在涉及补体激活的急性或慢性疾病中，通常需要有系统性炎症的标志物。现有的监测补体激活的方法比较繁琐，灵敏度也较低；其中包括溶血测定法（CH50）、液相末端复合物（C5b-C9）的定量以及沉淀凝胶技术（如 C3d）中补体片段产物的定量。我们开发了一种能够检测液相 C3c 的小鼠单克隆抗体（mAb），而不会受到其他由补体成分 C3 产生的产物的干扰。该 C3c 特异性 mAb 在不同的 ELISA 组合中进行了测试，包括各种类型的体外激活血清，以及因子 I 缺乏症患者的血浆或血清样本。该 mAb 的特异性通过免疫沉淀技术和免疫亲和层析后洗脱片段的分析进行了评估。该 C3c mAb 被证实是 C3c 特异性的，因为它与天然（未裂解）C3、C3b、iC3b 或 C3d 没有交叉反应。此外，在因子 I 缺乏症的血清或血浆中也没有观察到 C3 片段的显著反应。该抗体为生成强大的 ELISA 奠定了基础，该 ELISA 可快速可靠地评估补体激活和消耗作为炎症过程的标志物。我们在 100 名健康的丹麦献血者中建立了 C3c 血浆范围，平均值为 3.47μg/ml，范围为 2.12-4.92μg/ml。我们认为，在评估炎症过程中体内补体活性时，这种抗体可能具有潜在价值。
