肝癌细胞单克隆抗体磁珠对人肝癌、胃癌细胞株特异性富集的实验研究

摘要

目的 探讨肝癌细胞单克隆抗体(4-E9)免疫磁珠与上皮黏附分子(Epcam)磁珠的联合应用能否提高免疫磁珠对人乳腺癌细胞(MCF-7)、人肝癌细胞(BEL-7402)、人胃癌细胞(BGC-823)的富集效率.方法 通过生物素、链霉亲和素将制备的单克隆抗体与磁珠连接,制备成免疫磁珠.通过制备的两种免疫磁珠及两种免疫磁珠的联合应用分别对 MCF-7, BEL-7402和BGC-823细胞进行富集率检测.结果 4-E9免疫磁珠的包被率为57.8%,Epcam 免疫磁珠的包被率为65.8%.4-E9免疫磁珠对 MCF-7,BGC-823和 BEL-7402细胞富集率分别为(44±5.33)%,(71±11.33)%和(78.3± 8.46)%,Epcam免疫磁珠对 BGC-823,BEL-7402和 MCF-7细胞富集率分别为(55.5 ± 8.67)%,(78.88 ± 13.11)%和(84.31±6.83)%,两种免疫磁珠联合对 BGC-823,BEL-7402和 MCF-7细胞富集率分别为(80.4±8.33)%,(85.125± 6.77)%和(93.23±4.33)%,联合磁珠组较4-E9免疫磁珠对 BGC-823,BEL-7402,MCF-7细胞富集率差异均有统计学意义(P值分别为0.03,0.03,0.04).联合磁珠组较 Epcam免疫磁珠对BGC-823,BEL-7402,MCF-7细胞富集率差异均有统计学意义(P分别为0.04,0.03,0.04).结论 两种免疫磁珠联合使用能显著提高Epcam免疫磁珠对BEL-7402,BGC-823及 MCF-7细胞富集率,4-E9抗体对循环肿瘤细胞的富集可能具有一定的临床应用价值.%Objective To investigate whether the combination of 4-E9 immunomagnetic beads and epithelial adhesion mole-cules(Epacm)beads can enhance the enrichment efficiency of MCF-7,BEL-7402 and BGC-823 cells.Methods A mono-clonal antibody was prepared and ligated with magnetic bead by a biotin and a streptavidin to prepare an immunomagnetic beads.The enrichment rate of MCF-7,BEL-7402 and BGC-823 cells was detected by the combination of two kinds of immu-nomagnetic beads and two kinds of immunomagnetic beads.Results The encapsulation rate of 4-E9 immunomagnetic beads was 57.8%,and the encapsulation rate of Epcam immunomagnetic beads was 65.8%.4-E9 immunomagnetic beads on MCF-7,BGC-823 and BEL-7402 cell capture rate was(44±5.33)%,(71±11.33)% and(78.3±8.46)% respectively.Epcam immunomagnetic beads on BGC-823,BEL-7402 and MCF-7 cell capture rate was(55.5±8.67)%,(78.88±13.11)% and (84.31±6.83)% respectively.The combination of two kinds of immunomagnetic beads on BGC-823,BEL-7402 and MCF-7cell capture rate was(80.4±8.33)%,(85.125±6.77)% and(93.23±4.33)% respectively.Joint beads group compared with 4-E9 immunomagnetic beads on BGC-823,BEL-7402 and MCF-7 cell enrichment rate were statistically significant(P=0.03,0.03,0.04),and joint beads group compared with 4-E9 immunomagnetic beads on BGC-823,BEL-7402 and MCF-7 cell enrichment rate were statistically signific(P=0.04,0.03,0.04).Conclusion The combination of two kinds of immunomag-netic beads can significantly improve the enrichment efficiency of Epcam immunomagnetic beads on BEL-7402,BGC-823 and MCF-7 cells.4-E9 antibody enrichment of circulating tumor cells may have some clinical value.