CD59天然抗原的制备及纯化

目的 构建人matureCD59-mFC—pBOS真核表达载体，转染COSa细胞进行瞬时表达，对表达产物进行鉴定及纯化。方法在人Jurkat细胞中提取细胞总RNA，运用RT—PCR方法扩增matureCD59基因，定向克隆入带有mFC标签的真核表达载体pBOS中，大提质粒后通过电转仪转染COSa细胞，ELISA及特异抗体检测CD59的表达，并大量收集细胞上清用ProteinA进行纯化。结果构建r重组载体matureCD59．mFC—pBOS，瞬时转染COSa细胞，初步纯化得到天然表达的CD59抗原。结论带有mFC标签的CD59天然抗原的得到为制备抗人CD59单克隆抗体开辟了一条新的途径。

Preparation and Purification of CD59 Natural Antigen

Objective To construct eukaryotic vector expressing human mature CD59-mFC- pBOS, transfect COSa cells and obtain CD59 natural antigen. Methods The RNA was extracted from Jurkat and mature CD59 gene was amplified by RT-PCR. The CD59 gene was cloned into pBOS eukaryotic expression vector, named as matureCD59-mFC-pBOS. The recombinant plasmid was transfected transiently into COSa cells by electroporator. Superuatant CD59 was identified by ELISA and purified with Protein A. Results The eukaryotic expression vector CD59-mFC-pBOS was constructed. CD59 natural antigen was obtained through Protein A purification. Conclusion The purified CD59 natural antigen with mFC tag may be used for preparation of monoclonal antibodies against human CD59.