动物医学进展Issue(12)：5-9,5.
角蛋白19片段单克隆抗体的制备及鉴定
Preparation and Identification of Monoclonal Antibodies Against Cytokeratin 19 Fragment
摘要
Abstract
Our aim was to prepare monoclonal antibodies against cytokeratin 19 fragments (CYFRA21-1). Antibody titer of immunized mice was detected by indirect ELISA method.Monoclonal cell strains were obtained by cell fusion and screen.Antibody from ascites was produced by peritoneal injection the mono-clonal cell strains,and was purified by ammonium sulfate precipitation method and protein A chromatogra-phy.Antibody purity was detected by SDS-PAGE gel electrophoresis,and antibody concentration was checked by UV spectrophotometry.The antibody subtype kit was used to identify the subtypes of mono-clonal antibodies.The sensitivity of monoclonal antibody was measured by blocking ELISA method (IC50 ).The results showed that 4 monoclonal cell stains were produced after cell fusion and screen.Anti-body titer of 4 strain cells,which were 2F9,3H5,7C3,6F11,respectively were 2.56×10 5 ,5.12×10 5 ,5.12 ×105 ,2.56×105 ,and the range of concentration of 4 strain cells was 12.43 mg/mL-16.00 mg/mL,which was IgG1 antibody subtype.The inhibitory concentration 50 (IC50 )of antibody was in the range of 1.03 ng/mL-1.53 ng/mL.In a word,we obtained 4 monoclonal antibodies with high-titer against CY-FRA21-1 to establish CYFRA21-1 detection kit.关键词
角蛋白19片段(CYFRA21-1)/单克隆抗体/IgG/鉴定Key words
cytokeratin 19 fragment/monoclonal antibody/IgG/identification分类
生物科学引用本文复制引用
张慧茹,翟晋豫,刘珍,王雪琴,孟素香..角蛋白19片段单克隆抗体的制备及鉴定[J].动物医学进展,2014,(12):5-9,5.基金项目
河南省攻关项目（132102310169）；河南省重点攻关项目 （）
