【摘要】 采用RT PCR技术,从 1株稳定分泌抗人L-PGDS单克隆抗体的鼠杂交瘤细胞中扩增抗体的可变区基因,得 2个Vκ和 1个VH.序列经Igblast比对分析,其中 1个Vκ为鼠骨髓瘤细胞系内固有的无功能基因;另一Vκ和VH具备鼠抗体可变区特征,为抗体功能基因.经双酶切,将抗体可变区功能基因分别与表达载体pAG4622、pAH4604中的人IgG相应恒定区相连,构建成抗人L-PGDS的人 鼠嵌合抗体基因.更多还原

【Abstract】 The variable region genes of L-PGDS McAb are amplified from hybridoma cells by RT-PCR. After sequenced, one VH and two Vκ are obtained. But one of the Vκ is an aberrant Vκ transcript, derived from myeloma cell line. The other VH and Vκ are functional genes, for they are highly homologous to the variable region genes of murine immunoglobulin. Making use of enzymatic digestions, the VH and Vκ are joined to pAG4622 and pAH4604 to construct L-PGDS human-mouse chimeric antibody genes.更多还原