Shima N, Higashio K, Ogaki H, Okabe K
Life Science Research Institute, Snow Brand Milk Products Co., Ltd. Tochigi, Japan.
Gastroenterol Jpn. 1991 Aug;26(4):477-82. doi: 10.1007/BF02782817.
For determination of fibroblast-derived tumor cytotoxic factor, F-TCF (human hepatocyte growth factor/hHGF), sensitive two-step sandwich enzyme-linked immunosorbent assay (ELISA) employing monoclonal antibodies was developed. Microplates were coated with monoclonal antibody (P1C8) and bound F-TCF was quantitated with the second monoclonal antibody (P2D6) linked to peroxidase. The standard curve for F-TCF was found to be linear in the range of 0.16 to 10 ng of F-TCF per ml. The assay was specific for F-TCF but not for plasminogen. The assay can be used for determination of F-TCF antigen in both human plasma and serum. The variation of absorbance was little in duplicate samples. Recoveries of exogenous F-TCF added to serum or plasma samples showed theoretical values. F-TCF antigen levels in 21 healthy volunteers was found to be 0.56 +/- 0.43 ng/ml. In contrast, mean F-TCF levels in patients with liver diseases were all higher than those of healthy subjects. This ELISA system has the advantage of using a sensitive and reproducible set of monoclonal antibodies, and is a useful method for monitoring F-TCF levels in patients with liver diseases.
为了测定成纤维细胞衍生的肿瘤细胞毒性因子F-TCF（人肝细胞生长因子/hHGF），开发了一种采用单克隆抗体的灵敏的两步夹心酶联免疫吸附测定法（ELISA）。微孔板用单克隆抗体（P1C8）包被，结合的F-TCF用与过氧化物酶相连的第二种单克隆抗体（P2D6）进行定量。发现F-TCF的标准曲线在每毫升0.16至10 ng F-TCF范围内呈线性。该测定法对F-TCF具有特异性，对纤溶酶原则无特异性。该测定法可用于测定人血浆和血清中的F-TCF抗原。重复样品的吸光度变化很小。添加到血清或血浆样品中的外源性F-TCF的回收率显示为理论值。发现21名健康志愿者的F-TCF抗原水平为0.56±0.43 ng/ml。相比之下，肝病患者的平均F-TCF水平均高于健康受试者。这种ELISA系统具有使用一组灵敏且可重复的单克隆抗体的优点，是监测肝病患者F-TCF水平的一种有用方法。
