分泌抗CD71人-鼠嵌合抗体的转染瘤细胞分泌抗体的特异性与稳定性的研究

The study of specificity and stability of an anti-CD71 mouse/human chimeric antibody(Dd2C)-secreted transfectomas keeping in liquid nitrigen

摘要: 目的　研究在液氮中冻存 2年的分泌抗CD71人鼠嵌合抗体的转染瘤细胞分泌抗体的特异性、稳定性及抗体产量。方法　利用人IgG和鼠IgG作为浓度标准, 绘制浓度曲线, 比较研究转染瘤细胞培养上清的抗体分泌量。腹腔接种转染瘤细胞诱导裸鼠产生腹水抗体, 经离子交换法纯化, 电泳鉴定纯化嵌合抗体。结果　 1× 10⁵个 5ml转染瘤细胞培养 5天的上清中, 嵌合抗体分泌量为 (0 .5～ 5 ) μg ml。Balb c裸鼠腹腔接种转染瘤细胞后, 每只裸鼠腹水量在 (3～ 5 )ml,腹水抗体经纯化, 抗体产量约为 (1～ 2 )mg (ml腹水 )。经离子交换法纯化, 电泳鉴定, 在分子量 5 5kDa和 2 5kDa处, 可见有抗体IgG蛋白质的重链和轻链的染色条带。结论　由我们制备的在液氮中冻存 2年的分泌抗CD71人鼠嵌合抗体 (D2C)的转染瘤细胞体外生长良好, 抗体分泌稳定, 产量高, 特异性强。

Abstract: Objective To study the specificity and stability of an anti CD71 mouse/human chimeric antibody (D2C) secreted transfectomas keeping in liquid nitrogen for tow years. Methods Analysis of the chimeric Ab production in culture supernatant was made by the standard concentration curve method with ELISA. The antibody (D2C) was purified by DEAE Sephredax A50 ion exchange chromatography from ascites and was confirmed by SDS PAGE . Results The production of the chimeric Ab(D2C) in the culture supernatant wa...