Stable production of recombinant human sperm immobilizing antibody and its Fab fragment using cDNA expression vectors

利用cDNA表达载体稳定生产重组人精子固定化抗体及其Fab片段

• 批准号: 02671043
• 负责人: YAMASAKI Noriyuki
• 金额: $ 1.47万
• 依托单位: Hyogo Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02671043/
• 关键词: sperm immobilizing antibody; recombinant antibody; cDNA expression; gene transfection; BPV vector

Sperm immobilization antibodies present in sterile women are one of theprincipal immunological causes of infertility. To identify the antigenseliciting these antibodies, we have constructed cell lines that stably secrete recombinant human sperm-immobilizing antibody using cDNA expression vectors. For production of the antibody, cDNAs of the heavy and light chain immunoglobulingenes were cloned from the class-switched (from IgM to IgG) human sperm-immobilizing antibody En46 and inserted into modified bovine papilloma virus-based cDNAexpression vectors BCMGSHyg and BCMGSNeo, respectively. These constructs were sequentially transfected into the mouse myeloma cell line X63Ag8.653 by electro-poration. The established transfectants produced recombinant antibody that retained human sperm-immobilizing activity in non-selective medium for more thanone month. The use of this genetically engineered antibody should contribute to the understanding and treatment of immunological infertility and to the development of a contraceptive vaccine.

不育妇女体内存在的精子固定抗体是导致不育的主要免疫学原因之一。为了鉴定筛选这些抗体的抗原，我们利用cDNA表达载体构建了能够稳定分泌重组人精子固定化抗体的细胞系。为制备该抗体，从切换类（从IgM到IgG）的人精子固定化抗体En46中克隆出重链和轻链免疫球蛋白的cdna，分别插入到改良的牛乳头瘤病毒cdna表达载体BCMGSHyg和BCMGSNeo中。将这些构建体依次电穿孔转染到小鼠骨髓瘤细胞系X63Ag8.653中。所建立的转染物产生的重组抗体在非选择性培养基中保持人精子固定化活性超过一个月。这种基因工程抗体的使用应有助于了解和治疗免疫性不孕症，并有助于研制避孕疫苗。

项目成果

Hideaki Sawai, Noriyuki Yamasaki, Shinji Komori, Koji Koyama, Shinzo Isojima: "Stable Production of Recombinant Human Sperm Immobilizing Antibody using cDNA Expression Vectors"

Hideaki Sawai、Noriyuki Yamasaki、Shinji Komori、Koji Koyama、Shinzo Isojima：“使用 cDNA 表达载体稳定生产重组人精子固定化抗体”

K.ベクタ"Stable Production of Recombinant Human Sperm Immobilizing Antibody using cDNA Expression Vectors"

K. 载体“使用 cDNA 表达载体稳定生产重组人精子固定化抗体”