抗3α-HSD单克隆抗体制备方法的初步研究

摘要

Hybridoma technology was used to prepare 3α-HSD monoclonal antibody. Using 3α-HSD to express and pu-rify as antigen to immune Balb/c mice, the sensitization of B lymphocytes that against the antigen was gained. The hybridoma cell lines is got by fusing the B lymphocyte cell with SP2/0. Screen positive hole in ELISA, then the cell lines is cloned and the last is to screen the cell lines stabling to secrete 3α-HSD monoclonal antibody hybridoma. Al-though cell lines which could secrete monoclonal antibody hybridoma against 3α-HSD have been gotten, many special phenomenons were discovered during the preparation, such as qing on hybridoma with single ascites titer resistance is too low, but high serum titer. In the process of preparing single resistance, a preliminary analysis for against 3α-HSD monoclonal antibody preparation were made and technical support would be provided.%为了利用杂交瘤技术制备抗3α-HSD单克隆抗体，用表达纯化的3α-HSD作为抗原免疫Balb/c小鼠，获得针对该抗原致敏的B淋巴细胞，并用获得的B淋巴细胞与SP2/0进行细胞融合，得到杂交瘤细胞株，在ELISA间接法筛选出阳性孔的基础上，进行亚克隆，筛选出能稳定分泌抗3α-HSD单克隆抗体的杂交瘤细胞株。结果得到能分泌抗3α-HSD单克隆抗体的杂交瘤细胞株。制备过程中发现很多特殊的现象，如杂交瘤细胞上清与单抗腹水效价过低，但多抗血清效价较高等。文章对在单抗的制备过程中遇到的问题做了初步分析，为之后制备抗3α-HSD单克隆抗体的研究提供技术支持。