Raji cell line-specific Fab phage antibody library and screening.doc

 PAGE \* MERGEFORMAT 22 Raji cell line-specific Fab phage antibody library and screening Author: Xie Xiaofang, Shen Yongmei, Xiao-Chun Yang, Dong Ning Zheng, Xia Bai, Shi Yi Zhen [Keywords:] B-cell lymphoma;, Fab;, phage antibody libraries, Construction and screening of the specific Fab phage antibody library against Raji cell strain [Abstract] AIM: To construct and screen the specific Fab phage antibody library against human Raji cell strain in Blymphoma. METHODS: BALB / c mice were immunized with Raji cells, and the antibody light chain к genes and heavy chain genes Fd from the spleen cells were amplified by RTPCR. After restrictive digestion with Sac I / Xba I and Xho I / Spe I, the light chain к genes and heavy chain genes Fd were inserted into the phagemid vector pComb3HSS successively and then electroporated into E.coli XL1Blue. The specific Fab phage antibody library against Raji cell strain in human Blymphoma was constructed by infection of helper phage VCSM13. The specific antibodies against Raji cells were obtained after selected with Raji cells. The binding activity with antigens was identified by ELISA and the positive clones were sequenced. RESULTS: The Fab phage antibody library with 2.18 * 107 volume was constructed and eight positive clones which specifically recognized Raji cell strain were isolated. Sequence analysis of the two positive clones showed that the variable heavy domains (VH) and variable light domains (VL) were highly homologous with the registered murine Ig heavy chain V region sequences and kappa light chain sequences, respectively. CONCLUSION: Fab phage antibody library was successfully constructed and specific antibodies against membrane antigens in Raji cells were obtained, which will provide an experimental foundation for the further investigation of Blymphoma immunotherapy. [Keywords] Blymphoma; Fab; phage antibody library [Abstract] Objective: To construct for the B-cell lymphoma cell line Raji phage antibody library