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Cloning of heavy chain Fd gene of an anti-human-γ seminoprotein monoclonal antibody and its expression in HeLa cells

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Author:
No author available
Journal Title:
JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
Issue:
17
DOI:
10.3321/j.issn:1000-2790.2002.17.005
Key Word:
人精浆蛋白;单克隆抗体;免疫球蛋白重链Fd段;基因表达

Abstract: 目的获得鼠抗人精浆蛋白mAb重链Fd段基因,并将其转染到HeLa细胞进行表达. 方法从分泌抗人精浆蛋白mAb的杂交瘤细胞系E4B7中提取总RNA,用RT-PCR法获取重链Fd段cDNA. 将其克隆入真核表达载体pcDNA3,用脂质体转染法转染HeLa细胞,并用免疫荧光染色方法检测重链Fd段的表达. 结果从分泌抗人精浆蛋白的鼠mAb杂交瘤细胞系E4B7中克隆出了重链Fd段基因,序列测定表明,VH与鼠免疫球蛋白MUSIGHAEI同源性最高. 将含重链Fd段基因的真核表达载体pcDNA3- E4B7 Fd中转染HeLa细胞后,在HeLa细胞中检测到了重链Fd段的表达. 结论获得了序列正确的重链Fd段基因,它在HeLa细胞中可以成功地表达,为进一步构建和表达Fab奠定基础.

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