Stable expression of targeting complement inhibitor CR2-CD59 in Chinese hamster ovary cells

郭彦; 寇志华; 孙世惠; 张传福; 赵光宇; 于虹; 宋宏彬; 乔飞; 陈万荣; 周育森

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Stable expression of targeting complement inhibitor CR2-CD59 in Chinese hamster ovary cells

更新时间：2026-01-09

- Stable expression of targeting complement inhibitor CR2-CD59 in Chinese hamster ovary cells
- Journal of Army Medical University Vol. 32, Issue 9, Pages: 887-890(2010)
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  CLC： R322.65;R392.11;{R392.13}
- Published：2010
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郭彦,寇志华,孙世惠,张传福,赵光宇,于虹,宋宏彬,乔飞,陈万荣,周育森.人源靶向补体抑制物CR2-CD59在中国仓鼠卵巢细胞中的稳定表达[J].陆军军医大学学报(原第三军医大学学报),2010,32(09):887-890.

　Guo Yan,Kou Zhihua,Sun Shihui,Zhang Chuanfu,Zhao Guangyu,Yu Hong,Song Hongbin,Qiao Fei,Chen Wanrong,Zhou Yusen.Stable expression of targeting complement inhibitor CR2-CD59 in Chinese hamster ovary cells[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(09):887-890.
郭彦,寇志华,孙世惠,张传福,赵光宇,于虹,宋宏彬,乔飞,陈万荣,周育森.人源靶向补体抑制物CR2-CD59在中国仓鼠卵巢细胞中的稳定表达[J].陆军军医大学学报(原第三军医大学学报),2010,32(09):887-890. DOI：

　Guo Yan,Kou Zhihua,Sun Shihui,Zhang Chuanfu,Zhao Guangyu,Yu Hong,Song Hongbin,Qiao Fei,Chen Wanrong,Zhou Yusen.Stable expression of targeting complement inhibitor CR2-CD59 in Chinese hamster ovary cells[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(09):887-890. DOI：

摘要

目的构建人源靶向补体抑制物CR2-CD59，并筛选中国仓鼠卵巢细胞（Chinese hamster ovary cell，CHO）高效表达细胞株。方法运用FuGENE 6转染试剂，将含有人CR2-CD59的重组PEE14.1质粒转入CHO细胞，蛋氨酸亚氨基代砜（MSX）筛选出阳性克隆，并利用无血清培养基对CHO细胞表达株进行培养获得重组蛋白，以ELISA、SDS-PAGE和Western blot对表达蛋白进行鉴定。结果成功构建PEE14.1-CR2-CD59重组质粒，获得CHO细胞稳定表达株。SDS-PAGE结果表明，重组蛋白CR2-CD59的相对分子质量同预期结果一致。ELISA和Western blot鉴定重组蛋白CR2-CD59可与CR2、CD59多克隆抗体特异性结合。且与含血清培养基相比，无血清培养基能明显提高CHO细胞的蛋白表达量（P0.05）。结论在CHO细胞中成功表达人源靶向补体抑制物CR2-CD59。

Abstract

Objective To obtain Chinese hamster ovary (CHO) cell lines that stably express a targeting complement inhibitor CR2-CD59. Methods The recombinant plasmid PEE14.1-CR2-CD59 was constructed by cloning the DNA fragment CR2-CD59 into plasmid PEE14.1

and the obtained plasmid was transfected into CHO cells by FuGENE 6. The clones with stable high expression of target fragment were selected by methionine sulfoximine (MSX)

the expression of CR2-CD59 was analyzed by ELISA

SDS-PAGE and Western blotting analysis. Results Several stable expression clones were obtained

and CR2-CD59 was highly expressed in the secret form in CHO cells. SDS-PAGE analysis showed that the molecular weight of the recombined protein CR2-CD59 was consistent with the predicted one. ELISA and Western blotting results revealed that the CR2-CD59 could react with both anti-human CR2 and anti-human CD59 polyclonal antibodies. Compared with serum-containing medium

the protein was highly expressed in serum-free medium (P0.05). Conclusion Targeting complement inhibitor CR2-CD59 is successfully expressed in CHO cells.

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