睾酮单克隆杂交瘤细胞株的筛选及性能鉴定

摘要

为建立大批量样品中睾酮残留的检测方法,采用细胞融合技术、间接ELISA和间接竞争ELISA (icELISA)等方法,对睾酮单克隆杂交瘤细胞株进行了筛选及性能鉴定.结果表明:筛选出5株(T2C3,T2D9,T3A5,T4B1、T4E8)杂交瘤细胞,抗体亚类均为IgG1型,具k轻链,亲和常数为4.38×108～5.24×109 L/mol;腹水纯化后的抗体效价和灵敏度(IG50)分别为0.64×105～2.56×105和1.58～2.92ng/mL.以T2D9细胞株建立icELISA标准曲线,其线性范围为0.06～63.5 ng/mL,检测限和IC50值分别为0.04和1.55 ng/mL.%To establish the testing methods of testosterone residues in multitudinous samples, the selection and performance appraisal of monoclonal hybridoma cell lines of testosterone were conducted through the methods of cell fusion technology, indirect ELISA and indirect competitive ELISA (icELISA). The results showed that the five hybridoma cell lines named T2 C3, T2D9, T3 A5, T4 B1 and T4E8 were screened out. The corresponding mAbs were of the IgG1 isotype with a k light chain, and the antibody affinity of all mAbs were between 4. 38 × 108 and 5. 24 × 109 L/mol. The titer and IC50 values of purified ascites were in the range of 0. 64×105~2. 56×105 and 1. 58~2. 92 ng/mL, respectively. Based on the T2D9 hybridoma, an icELISA standard curve was developed, and the dynamic range was from 0. 06 to 63. 5 ng/mL, with a detection limit and IC50 values of 0. 04 ng/mL and 1. 55 ng/mL, respectively.