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目的为了在CHO-dhfr 细胞中高效表达有活性的抗人CEA鼠-人嵌合抗体。方法构建抗CEA嵌合抗体的真核高效表达载体,转染CHO-dhfr-细胞,通过加入 MTX进行基因扩增表达,获得高产细胞株。采用Western bolt、ELISA 、体内放射免疫实验等方法鉴定所表达的嵌合抗体的生物学特性。结果获得产量可达60 μg/ml的细胞株。Western blot、 ELISA、体内放射免疫实验证明所表达的嵌合抗体的确含人抗体的恒定区,并特异地与 CEA结合。结论成功地在真核细胞CHO-dh fr-中高效表达了抗CEA嵌合抗体,并具有良好的生物活性。
OBJECTIVE To efficiently express active anti-human CEA murine-human chimeric antibodies in CHO-dhfr cells. Methods Eukaryotic expression vector of anti-CEA chimeric antibody was constructed and transfected into CHO-dhfr- cells. MTX was used for gene amplification and expression, and the high-yielding cell line was obtained. Western blot, ELISA and in vivo radioimmunoassay were used to identify the biological characteristics of the expressed chimeric antibodies. As a result, cell lines producing a yield of 60 μg / ml were obtained. Western blot, ELISA, in vivo radioimmunoassay showed that the expressed chimeric antibody did contain the constant region of human antibody and specifically bound to CEA. Conclusion The anti-CEA chimeric antibody was successfully expressed in eukaryotic CHO-dh fr- and has good bioactivity.