抗人胰腺癌单克隆抗体的制备及其特性的研究

The Production of the Monoclonal Antibody to Human Pancreatic Cancer and the Research of its Features

本实验采用常规细胞融合方法，在ＰＥＧ作用下，成功地制备了五株能稳定分泌抗人胰腺癌单克隆抗体的杂交瘤。以间接型ＥＬＩＳＡ方法和ＡＢＣ技术进行抗体检测，有限稀释法进行克隆。用羟基磷灰石法纯化单克隆抗体（ＭＡｂ）．所得ＭＡｂＢ６的免疫球蛋白类型为ＩｇＧ１，它在培养上清及小鼠腹水中的效价分别为１：１６和１：２６００。ＭＡｂＢ６具有较好的特异性，其所针对的抗原可能为一种胰腺癌细胞胞膜抗原，是一种肿瘤相关性抗原。羟基磷灰石方法是一种比较理想的单克隆抗体提纯方法。

Using routine cellular hybridization method,we made five strain hybridoma cells which could secrect monoclonal antibodies(MAbs) to human pancreatic cancer. ELISA and ABC method were employed to screen. The desired hybridomas were cloned by limiting dilution. MAbs were purified by hydroxylapatite chromatography. Subclass of MAb B6, one of the five MAbs,was IgG1. The titres of MAb B6 in culture and ascitic fluid were 1: 16 and 1: 2600 respectively. MAb B6 possessed considerable specificity. The antigen that MAb B6 identified was a membranous antigen expressing mainly in the cell membrances of pancreatic cancer. It was a tumor-relative antigen, not a tumor-specific antigen. Hydroxylapatite chromatography was an ideal method for antibody purification.