抗人PCIA1单克隆抗体的制备及其生物特征的研究

Production of Monoclonal Antibody Against Human PCIA1 and Study on Its Characteristics

目的:筛选制备抗人PCIA1的单克隆抗体,鉴定其生物学特征。方法:采用原核表达并纯化的PCIA1蛋白作为抗原,采用杂交瘤技术制备抗人PCIA1单克隆抗体的杂交瘤,用ELISA进行筛选和鉴定其亚类,并用细胞扒片免疫化学方法检测其反应性。结果:获得了32株分泌抗人PCIA1的McAb杂交瘤细胞,McAb-1A4的染色体呈双亲特点,该McAb与多种人肿瘤细胞株和正常肝细胞株有反应。结论:McAb-1A4可用于PCIA1的细胞内定位研究。

Objective： To prepare monoclonal antibody （McAb） against human PCIA1 protein and study its biological activity. Methods： As an antigen recombinant PCIA1 protein was used for immunizing Balb/c mice. McAbs against PCIA1 protein was prepared by hybridoma technique, ELISA was used to select hybridomas and identify its immunoglobin subclass, Attach cell model also was used to identify where PCIA1 existed in cells. Results： Thirty - two hybridomas which secreted McAb against PCIA1 were obtained, The chromosomes of McAb - 1A4 showed the characteristics of their parents. The immunoglobulin subclass of McAb - 1A4 was IgG1. This McAb presented reactions to all tumor cell lines and hepatoceUtdar normal cell line adopted. Conclusion： McAb- 1A4 is able to be used to localize the site of PCIA1 in cells.