Human source of anti-hepatoma single-chain antibody library and preliminary identification of.docVIP

 PAGE \* MERGEFORMAT 18 Human source of anti-hepatoma single-chain antibody library and preliminary identification of 【Abstract】 Objective To construct humanized single-chain antibody against hepatocellular carcinoma gene phage display library. Methods using phage display technology to construct genomic library enriched after panning, after screening, using ELISA means of checking the antigen binding activity. Results from 30 phage clones were screened to 8 with a hepatoma cell line SMMC7721 binding activity of the positive clones. Conclusions obtained from peripheral blood lymphocytes variable region genes, the use of phage antibody library technology to prepare humanized single-chain antibody against hepatocellular carcinoma strategy is feasible. Keywords: phage display technology a single chain antibody liver 0 Introduction Liver cancer in China a common malignancy, current treatment initiatives in the hepatectomy-based comprehensive treatment. But for the early diagnosis of liver cancer, treatment effectiveness has not been satisfactory [1]. Single-chain antibody (ScFv) is to use genetic engineering method to antibody heavy chain variable region (VH) and light chain variable region (VL) via a connecting peptide (Linker) connections made of recombinant proteins is to maintain the parent antibody antigen affinity and activity and specificity of the smallest functional antibody fragment [2]. Construction of anti-phage antibody library and the preparation of HCC-specific single-chain antibody with a larger value of basic research and broad prospect for clinical application [3]. This experiment is the use of phage display technology to construct a single-chain antibody against human liver cancer gene phage display library and screened from antibodies are antigen-binding activity. 1 Materials and methods 1.1 Materials Trizol reagent, cDNA Synthesis Kit and gel purification kit purchased from Gibco Company, E.co