抗人黑色素瘤嵌合抗体真核表达载体的构建与表达

目的：构建嵌合型抗人黑色素瘤抗体的真核表达载体，并实现真核表达。方法：从3株杂交瘤细胞中克隆得到鼠源单抗的可变区基因，插入含有抗人Tac抗原信号肽以及人免疫球蛋白κ轻链恒定区基因和γ1重链恒定区基因的真核表达载体pMH-CA中，转染293T细胞，进行嵌合抗体表达，并用RT-PCR、ELISA、Westem blot免疫印迹等对抗体表达鉴定。结果：成功构建了抗人黑色素瘤人，鼠嵌合抗体，并实现其真核表达。RT-PCR证实了该抗体在mRNA水平上的表达，ELISA和Westemblot证实了抗人黑色素瘤人，鼠嵌合抗体的表达。结论：成功表达了抗人黑色素瘤人-鼠嵌合抗体。

Construction and expression of mouse-human chimeric anti-human melanoma antibody

Objective:To construct eukaryotic expression vector for anti-human melanoma chimeric antibody,and express the antibody in eukaryotic cells.Methods:Variable regions of the murine antibody were cloned from the anti-human melanoma hybridoma cell line HB8459,HB8760,Mel3,and was inserted them into the chimeric antibody eukaryotic expression vector pMH-CA.The constructed vector was transfected into 293T,the chimeric antibody was expressed,as detected by RT-PCR,ELISA,and Western blot.Results:Eukaryotic expression vectors of the chimeric antibody against human melanoma were successfully constructed,and expressed in eukaryotic cells.The expression of chimeric gene was confirmed by RT-PCR,specificity of the chimeric antibody was proved by ELISA and Western blot.Conclusion:Anti-human melanoma chimeric antibody was expressed successfully.