Greffard A, Bourgarit J J, le Maho S, Lambré C R
Immunol Lett. 1987 Jun;15(2):145-51. doi: 10.1016/0165-2478(87)90046-0.
In order to measure the concentration of the human complement component C2 in various biological fluids, an enzyme linked immunosorbent assay (ELISA) was developed. This assay was highly sensitive and allowed to detect as few as 400 pg of C2 in a sample volume of 150 microliters (i.e. 2.6 ng/ml). This is a 10- to 15-fold increase in sensitivity with regard to the conventional hemolytic test. As assessed by an immunoblot analysis, our anti-C2 antiserum was able to detect native C2 as well as the cleavage fragments C2a and C2b generated upon complement activation through the classical pathway. Thus, complement activation involving the classical pathway can easily be evidenced by comparing functional (hemolytic) and immunochemical (ELISA) C2 assays which respectively do not and do reveal activated C2. When C2 was assayed in either normal human serum or bronchoalveolar fluids, in both ELISA and hemolytic tests, a highly significant correlation was observed between the two assays (P less than or equal to 0.01). The specific C2 activity (i.e. functional hemolytic activity/ng C2 assayed in ELISA) was higher in serum than in bronchoalveolar lavage fluids from both normal volunteers and patients with pulmonary diseases.
为了测量各种生物体液中人类补体成分C2的浓度，开发了一种酶联免疫吸附测定法（ELISA）。该测定法高度灵敏，在150微升的样本体积中能够检测低至400皮克的C2（即2.6纳克/毫升）。这比传统溶血试验的灵敏度提高了10至15倍。通过免疫印迹分析评估，我们的抗C2抗血清能够检测天然C2以及通过经典途径激活补体时产生的裂解片段C2a和C2b。因此，通过比较分别不能检测和能检测活化C2的功能（溶血）和免疫化学（ELISA）C2测定法，可以很容易地证明涉及经典途径的补体激活。当在正常人血清或支气管肺泡液中检测C2时，在ELISA和溶血试验中，两种测定法之间均观察到高度显著的相关性（P≤0.01）。正常志愿者和肺部疾病患者的血清中C2的比活性（即功能溶血活性/ELISA测定的纳克C2）高于支气管肺泡灌洗液。
