Application of PCR technology to build cells and screening of phage antibody library with vitiligo.docVIP

 PAGE \* MERGEFORMAT 17 Application of PCR technology to build cells and screening of phage antibody library with vitiligo Author: Ling, Tian Yan, Li Ying, high astronomical [Keywords:] Vitiligo; polymerase chain reaction; phage antibody library; autoantibodies Construction and screening of human vitiligo phage autoantibody library based on incell PCR [Abstract] AIM: To obtain in situ pairing of the variable region genes of the heavy and light chains of vitiligo patients lymphocytes, then construct and screen a human ScFv phage antibody library. METHODS: We amplified in situ pairing of the variable region genes of the immunoglobulin heavy and light chains using the incell PCR together with Crerecombination, constructed and panned a human ScFv phage antibody library, and screened it with pigment cell suspension, then detected the activity of positive clones. RESULTS: We constructed an antibody library of 3 * l06 in size, and obtained one specific autoantibody against melanocyte membrane antigens. CONCLUSION: Incell PCR together with human phage antibody library may be useful tools for the study of vitiligo antibody gene family and the VH / VL pairing that occurs during the autoimmune process. [Keywords] vitiligo; polymerase chain reaction; phage antibody library; autoantibodies [Abstract] Objective: To clone lymphocytes in patients with vitiligo in the original pair’s own antibody genes to construct phage antibody library and screening for the melanocyte-specific membrane antigen antibodies. Methods: The cell-PCR amplification of human immunoglobulin light chain and heavy chain variable region genes, Cre recombinase-mediated complete the light and heavy chain within a single cell connected to the body can be reflected in the original pairing information, a single-chain antibody variable region genes to construct phage antibody libraries, screening for melanin membrane antigen autoantibodies . Results: Construction of the completion of storage capacity of a