Dorer F E, Levine M, Skeggs L T, Lentz K E, Kahn J R
Hypertension. 1984 May-Jun;6(3):374-82. doi: 10.1161/01.hyp.6.3.374.
Spleen cells from mice immunized with partially purified hog kidney renin were fused with mouse myeloma cells to produce a stable monoclonal hybridoma cell line that synthesizes an antibody against renin. A single monoclonal antibody was chosen for study and has been produced in large quantity and purified by affinity chromatography on protein A-Sepharose. The antirenin, which belongs to the IgG1 subclass, exhibits anticatalytic activity against both hog and rabbit renin. An immunoaffinity column prepared from antibody coupled to Sepharose has been used in the purification of renin from hog kidney. Although renin is quantitatively adsorbed from solution, it can be eluted from the column under gentle conditions. The highly purified renin, with specific activity of 2122 Goldblatt Units/mg protein, exhibits both charge (pH 4.1 to 5.1) and size (38,000 to 42,700) heterogeneity. Hog kidney renin dissociates in the presence of sodium dodecyl sulfate (SDS) and mercaptoethanol to heavy and light chains with molecular weights of 33,700 and 5,800, respectively. In the presence of SDS, a small amount of a nw form of renin is observed with a molecular weight of 19,500 which retains activity on renaturation. The monoclonal antibody should be a useful tool for the study of the renin-angiotensin system and especially for the purification of renin. The hybridoma cell line used in this study (F-32 VIII C4) has been donated to the American Type Culture Collection.
用部分纯化的猪肾肾素免疫的小鼠脾细胞与小鼠骨髓瘤细胞融合，以产生稳定的单克隆杂交瘤细胞系，该细胞系可合成抗肾素抗体。选择了一种单克隆抗体进行研究，并大量生产，通过在蛋白A-琼脂糖凝胶上进行亲和层析进行纯化。该抗肾素属于IgG1亚类，对猪和兔肾素均表现出抗催化活性。由偶联到琼脂糖凝胶上的抗体制备的免疫亲和柱已用于从猪肾中纯化肾素。尽管肾素可从溶液中定量吸附，但可在温和条件下从柱上洗脱。高度纯化的肾素，比活性为2122 Goldblatt单位/毫克蛋白，表现出电荷（pH 4.1至5.1）和大小（38,000至42,700）的异质性。猪肾肾素在十二烷基硫酸钠（SDS）和巯基乙醇存在下解离为重链和轻链，分子量分别为33,700和5,800。在SDS存在下，观察到少量分子量为19,500的新形式肾素，其复性后仍保留活性。该单克隆抗体应是研究肾素-血管紧张素系统，特别是纯化肾素的有用工具。本研究中使用的杂交瘤细胞系（F-32 VIII C4）已捐赠给美国模式培养物保藏所。
