Liu Yong-mei, Feng Zhi-chun, Du Jiang, Wu Bing-yi, Tan Yue-er, Wang Xian-yan
Department of Pediatrics, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China.
Di Yi Jun Yi Da Xue Xue Bao. 2003 Aug;23(8):849-51.
To prepare monoclonal antibody (mAb) against human pulmonary surfactant-associated protein A (SP-A) and evaluate its specificities.
The hybridoma cell line secreting SP-A mAb was established to induce the production of SP-A ascites fluid in BALB/c mice immunized with SP-A. The ascites fluid was then collected and purified through (NH4)2SO4 salting-out procedure, and the titer and specificity of the purified monoclonal anti-SP-A antibodies were determined by Western blotting and indirect enzyme-linked immunosorbent assay, respectively.
Three hybridoma cell lines capable of specific antibody secretion were established, among which 2B6 produced the antibody of the highest titer, with the hybridoma cell chromosome exceeding 100. The antibody titers in the ascitic fluid and cell supernatant fluid were 1: 50,000 and 1: 10,000 respectively, and Western blotting displayed a reaction band representing a relative molecular mass of 31,000, which was produced by the monoclonal antibody in reaction to purified SP-A.
The antibodies of high purity and specificity against human SP-A have been successfully prepared.
制备抗人肺表面活性物质相关蛋白A（SP-A）的单克隆抗体（mAb）并评估其特异性。
建立分泌SP-A mAb的杂交瘤细胞系，在用SP-A免疫的BALB/c小鼠中诱导产生SP-A腹水。然后收集腹水并通过硫酸铵盐析法进行纯化，分别通过蛋白质印迹法和间接酶联免疫吸附测定法测定纯化的抗SP-A单克隆抗体的效价和特异性。
建立了3株能够分泌特异性抗体的杂交瘤细胞系，其中2B6产生的抗体效价最高，杂交瘤细胞染色体超过100条。腹水中和细胞上清液中的抗体效价分别为1:50000和1:10000，蛋白质印迹显示出一条代表相对分子质量为31000的反应带，这是单克隆抗体与纯化的SP-A反应产生的。
已成功制备出针对人SP-A的高纯度、特异性抗体。
