%0 Journal Article
%T Construction and preliminary identification of genetically engineered tetravalent antibodies against parathion<br>抗对硫磷基因工程新型抗体的制备及初步鉴定
%A Yaqi Qiao
%A Jiarong Pan
%A Wang Lei
%A Haiying Yu
%A Chen Min
%A Zhang Wei
%A Fukai Zhang
%A <br>乔亚奇
%A 潘家荣
%A 王磊
%A 于海英
%A 陈明
%A 张维
%A 张付凯
%J 微生物学报
%D 2010
%I 
%X Abstract: Objective]The objective is to improve the affinity of antibody against parathion with an aim to enhance the sensitivity of ELISA assay. Methods] We recombined anti-parathion single-chain variable fragment gene (scfv) and the core streptavidin gene(sa) to produce an anti-parathion single-chain variable fragment-core of streptavidin-biotin fusion gene (scfv-sa), and inserted the fusion gene (scfv-sa) into the vector pET28a (+) in E.coli BL21 for transformation by prokaryotic expression. The expression was analyzed with SDS-PAGE and Western blot. the affinity of the recombinant protein was measured by ELISA after purified by metal affinity chromatography using Ni-NTA. Results] The recombinant gene could express a fusion protein about 46 kDa, which formed a tetravalent domain, tetravalent antibody. ELISA results showed that the tetravalent antibody could bound to parathion specifically with the Affinity constant of 4.25 × 107 L / mol and titer of over 1:1×106. Conclusion] The recombining anti-parathion tetravalent antibody could react with parathion specifically with significantly more binding sites with the monoclonal antibody, based on which the detection sensitivity of ELISA would be improved.
%K parathion
%K core streptavidin
%K tetravalent antibodies
%K prokaryotic expression<br>关键词:
%K 对硫磷
%K 链霉亲和素
%K 四价抗体
%K 原核表达
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=E3990D156CE254FC87D130F084F487D2&yid=140ECF96957D60B2&vid=771152D1ADC1C0EB&iid=94C357A881DFC066&sid=2B6C525BCE31A7DA&eid=DEE640F0CDC9D495&journal_id=0001-6209&journal_name=微生物学报&referenced_num=1&reference_num=0