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

不育妇女体内存在的精子制动抗体是导致不育的主要免疫学原因之一。为了鉴定这些抗体的抗原选择性，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。为了生产抗体，从类别转换（从IgM到IgG）的人精子固定抗体En46中克隆重链和轻链免疫球蛋白基因的cDNA，并分别插入基于牛乳头状瘤病毒的cDNA表达载体BCMGSHyg和BCMGSNeo中。通过电穿孔将这些构建体依次转染到小鼠骨髓瘤细胞系X63Ag8.653中。建立的转染子产生的重组抗体在非选择性培养基中保持人精子固定活性超过一个月。这种基因工程抗体的使用应有助于理解和治疗免疫性不孕症和避孕疫苗的发展。

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.