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

不孕不育妇女中存在的精子固定抗体是不孕症的主要免疫学原因之一。为了鉴定这些抗体的抗原性，我们利用cDNA表达载体构建了稳定分泌重组人精子固定抗体的细胞系。为了制备抗体，从类转换的人精子固定抗体En46中克隆了重链和轻链免疫球蛋白基因，将其分别克隆到改良的牛乳头瘤病毒cDNA表达载体BCMGSHyg和BCMGSNeo中。用电穿孔法将其依次导入小鼠骨髓瘤细胞系X63Ag8.653。所建立的转基因细胞产生的重组抗体在非选择性介质中保持了一个多月的人精子固定化活性。这种基因工程抗体的使用应该有助于理解和治疗免疫性不孕症，并有助于开发避孕疫苗。

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.