中国免疫学杂志2013，Vol.29Issue(6)：649-652,4.DOI:10.3969/j.issn.1000-484X.2013.06.020
β2糖蛋白Ⅰ单克隆抗体的制备与鉴定
Preparation and identification of the monoclonal antibodies against β2-glycoprotein Ⅰ
摘要
Abstract
Objective:To produce and identify the monoclonal antibodies (MAbs) against β2-glycoprotein Ⅰ (β2 GP Ⅰ).Methods:The spleen cells from the BALB/c female mouse immunized with β2 GP Ⅰ were fused with SP2/0 mouse myeloma cells in PEG to generate hybridoma cells,which were screened by limited dilution.Then the hybridoma cells were injected to BALB/c mice to make ascites,in which the antibodies were produced abundantly,followed by purified with ammonium sulfate and protein G.Colchicine was used to identify the number of chromosome of hybridoma cells.The properties of the MAbs against β2 GP Ⅰ (the titer of the MAbs in the culture medium and ascites,the isotypes of the MAbs and the specificity of the MAbs) were determined by indirect ELISA,MAbs isotyping kit and Western blot,respectively.Results:One hybridoma cell line (β2),secreting MAbs against β2 GP Ⅰ,had been established.The number of chromosome of the hybridoma cells was between 99 and 108.Further studies found that the titer of the MAbs against β2GP Ⅰ in the culture medium and ascites was 1∶29 and 1∶215,respectively.The immunoglobulin of the MAbs was classified as IgG1/κ,with satisfactory specificity.Conclusion:The MAbs against β2GP Ⅰ were prepared and identified,which pave the way for the research in the mechanism of antiphospholipid syndrome.关键词
β2GP Ⅰ/杂交瘤细胞株/单克隆抗体/抗磷脂综合征Key words
β2GP Ⅰ/Hybridoma cell/Monoclonal antibody/Antiphospholipid syndrome分类
医药卫生引用本文复制引用
胥亚,许国莹,周红,解鸿翔,夏龙飞,刘敬敬,张晓蕾..β2糖蛋白Ⅰ单克隆抗体的制备与鉴定[J].中国免疫学杂志,2013,29(6):649-652,4.基金项目
本文受国家自然科学基金项目(No.30971301)资助 （No.30971301）
