CAJ | 학술논문

目的：研究解决半抗原分子单克隆抗体制备技术路径中遇到的在阳性杂交瘤细胞株筛选时无法排除载体蛋白间交叉反应影响的问题，以半抗原去甲肾上腺素（norepinephrine，NE）为例。方法：在NE完全抗原免疫小鼠实施细胞融合后，分别包被牛血清白蛋白（BSA）、卵清白蛋白（OVA）、BSA-NE、OVA-NE等4种不同抗原的酶标板平行检测细胞培养上清液；挑选BSA、OVA检测结果为阴性，BSA-NE、OVA-NE检测结果为阳性的孔内细胞进行克隆化筛选单克隆细胞。结果：本筛选方法可一次性从8板96孔板中筛选到13个符合要求的阳性孔，经3次克隆化后获得6株特异性强的杂交瘤细胞株。结论：本方法避免了载体蛋白间交叉反应对筛选的影响，改进了传统的单一指标筛选方法，筛选效率更高。
목적：연구해결반항원분자단극륭항체제비기술로경중우도적재양성잡교류세포주사선시무법배제재체단백간교차반응영향적문제，이반항원거갑신상선소（norepinephrine，NE）위례。방법：재NE완전항원면역소서실시세포융합후，분별포피우혈청백단백（BSA）、란청백단백（OVA）、BSA-NE、OVA-NE등4충불동항원적매표판평행검측세포배양상청액；도선BSA、OVA검측결과위음성，BSA-NE、OVA-NE검측결과위양성적공내세포진행극륭화사선단극륭세포。결과：본사선방법가일차성종8판96공판중사선도13개부합요구적양성공，경3차극륭화후획득6주특이성강적잡교류세포주。결론：본방법피면료재체단백간교차반응대사선적영향，개진료전통적단일지표사선방법，사선효솔경고。
Objective: Take norepinephrine(NE) as an example to establish an improved hybridoma screen mode which increasing specificity and efficiency of screening procedure when preparing for anti-hapten antibodies. Meth-ods: After cell fusion steps, four kinds of antigens, bovine serum albumin(BSA), ovalbumin(OVA), BSA-NE, OVA-NE, were coated separately on 96 wells ELISA plate and simultaneously test the supernatant of hybridoma culture. Those hybridomas wells with results of BSA-NE and OVA-NE positive reaction, but BSA and OVA nega-tive reaction were selected for a further cloning procedure to get monoclonal cell lines. Results: 13 satisfied mono-clonal hybridoma cells were obtained from 8 ELISA plates. After the cloning procedure 6 high specific monoclonal antibody secreting hybridomas cell lines acquired. Conclusion: This improved ELISA screening mode directly en-hanced the specificity towards hapten structure compared with currently commonly used method. This mode can suf-ficiently avoid interaction between protein carriers.