改进双抗夹心ELISA用于筛选单抗杂交瘤细胞株

本文利用改进双抗夹心ELISA筛选一种可测定血清样品中与血浆蛋白结合的蛋白质药物的单抗。用兔抗K102多抗经亲和层析纯化并包被后,加入用10%猴血浆PBS还原后的抗原K102,再加入杂交瘤细胞株上清液和HRP-羊抗鼠IgG、底物反应液筛选阳性单抗杂交瘤细胞株。结果用改进后的双抗夹心ELISA筛成功地选出6株能稳定分泌针对K102的单抗杂交瘤细胞株,分别命名为1C9、2D7、3E2、5F4、5F6、6B9;细胞株产生的单抗用ELISA法能检测出血清样品中的K102的浓度。提示改进的双抗夹心ELISA可用于针对与血浆蛋白结合的蛋白质药物的单抗杂交瘤细胞株的筛选。

Application of Improved Double-Antibody Sandwich Elisa to Screening Monclonal Antibody of Hybridoma Line

Objective: To screen monoclonal antibody against protein drugs bound with plasma protein in blood samples.Methods:After coated with rabbit anti-K102 polyclonal antibody purified by affinity chromatography,adding K102 antigen diluted with PBS containing 10% monkey plasma PBS and the supernatant of hybridoma and HRP-goat anti-mouse IgG are added to screen positive hybridoma lines which might produce monoclonal antibodies.Results:In use of the method of double-antibody sandwich ELISA screening,the six hybridoma lines are successfully screened to be capable of screening stably the antibody against K102,named as 1C9,2D7,3E2,5F4,5F6,6B9.The monoclonal antibodies could detect K102 concentration in the blood samples.Conclusion:The improved double-antibody sandwich ELISA can be used to screening hybridoma cell lines of monoclonal antibodies against biodrugs bound with plasma protein.