Fang Shi-Song, Zhao Yu-Mei, He Jian-Fan, Liu Tao, Liu Jianjun, Zhao Shujin
South China University of Technology, Guangzhou 510642, China.
Wei Sheng Yan Jiu. 2005 May;34(3):289-92.
To expression the phage single chain antibody against nucleoprotein of influenza virus A type in E. coli strain HB2151 by screening the positive clone from the phage antibody library against nucleoprotein, which will prepare for the construction of fast leak kit of Influenza virus A type.
The positive clone ratio in the phage antibody library was enriched by three-round continual solid panning, ELISA, SDS-PAGE and Western-blot methods were used to analyze the protein secreted into the supemants and periplasmic. Affinity chromatography was used to purified the protein expressed in periplasmic and the titer was also analyzed using ELISA method.
10 clones were screened from96 clones and among the 10, there were 3 stronger positive with OD450nm value of 0.469, 0.582 and 0.507, respectively. The phage single chain antibody against Influenza virus A type was primary expressed in periplasmic. The ELISA results were positive even if the single chain antibody purified by affinity chromatography was diluted 4096 folds.
Using phage antibody library technique, phage single antibody against nucleoprotein of influenza virus A type was preliminary expressed in E. coli.
通过从抗核蛋白噬菌体抗体库中筛选阳性克隆，在大肠杆菌HB2151菌株中表达抗甲型流感病毒核蛋白的噬菌体单链抗体，为甲型流感病毒快速检测试剂盒的构建做准备。
通过三轮连续固相淘选富集噬菌体抗体库中的阳性克隆比例，采用ELISA、SDS-PAGE和Western-blot方法分析培养上清液和周质中分泌的蛋白。采用亲和层析法纯化周质中表达的蛋白，并使用ELISA方法分析其效价。
从96个克隆中筛选出10个克隆，其中3个阳性较强，OD450nm值分别为0.469、0.582和0.507。抗甲型流感病毒的噬菌体单链抗体在周质中初步表达。经亲和层析纯化的单链抗体即使稀释4096倍，ELISA结果仍为阳性。
利用噬菌体抗体库技术，在大肠杆菌中初步表达了抗甲型流感病毒核蛋白的噬菌体单链抗体。
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