抗禽流感病毒H5N1分泌型IgA在中国仓鼠卵巢细胞中的表达  

Expression of anti-avian influenza virus H5N1 secretory IgA in Chinese hamster ovary cells

作　　者：李存[1] 张宝中[1] 安小平[1] 米志强[1] 刘大斌[1] 姜焕焕[1] 潘博[1] 王盛[1] 陈斌[1] 黄芬[1] 王娟[1] 王晓娜[1] 童贻刚[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《生物工程学报》2011年第2期219-225,共7页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.30872223);国家高技术研究发展计划(863计划)(No.2009AA02Z111)资助~~

摘　　要：分泌型IgA(SIgA)在机体的粘膜免疫中具有重要作用,在外分泌道中比单体IgA和IgG抗体具有更好的抗感染活性。为了表达抗禽流感病毒H5N1人-鼠嵌合分泌型IgA抗体,首先以本室先前构建的稳定表达IgA的中国仓鼠卵巢细胞(CHO)细胞系为基础,共转染分泌片和J链表达质粒,然后用抗生素Zeocin选择阳性克隆细胞,利用倍比稀释的方法筛选分泌SIgA的单克隆细胞,通过Western blotting分析培养上清中SIgA的表达情况。结果表明,在CHO细胞中成功表达了SIgA抗体,上述研究为研制分泌型IgA抗体制剂奠定了良好的基础。Secretory IgA（SIgA） antibodies in external secretions play an important role in mucosal immune response.Polymeric SIgA was advantageous over monomeric IgA（mIgA） and IgG in several aspects.To express secretory IgA antibody against H5N1 virus,we constructed the secretory component and immunoglobulin J expressing plasmids and co-transfected the plasmids into the Chinese hamster ovary cells（CHO） stably expressing immunoglobulin A.Then we used Zeocin to select the positive clone cells,monoclonal cells stably secreting SIgA was screened through fold dilution method at last.The SIgA antibody secreted from the CHO cells was confirmed by Western blotting,which demonstrated that we had got the complete SIgA molecular.The successful expression of this polymeric anti-H5N1 SIgA in CHO cells will contribute to the production of recombinant SIgA as a preventive agent for infectious disease control.

关 键 词：禽流感病毒H5N1 WESTERN BLOTTING SIgA抗体 

分 类 号：Q78[生物学—分子生物学]