Construction and screening of the specific Fab phage antibody library against Raji cell strain

    XIE Xiaoª²fang, SHEN Yongª²mei*, YANG Xiaoª²chun, DONG Ningª²zheng, BAI Xia, SHI Yiª²zhen

    Department of Clinical Laboratory, Second Affiliated Hospital of Soochow University, Suzhou 215004; Jiangsu Institution of Hematology, First Affiliated Hospital of Soochow University, Suzhou 215006, China

    [Abstract] AIM: To construct and screen the specific Fab phage antibody library against human Raji cell strain in Bª²lymphoma. METHODS: BALB/c mice were immunized with Raji cells, and the antibody light chain §Ü genes and heavy chain genes Fd from the spleen cells were amplified by RTª²PCR. After restrictive digestion with Sac I/Xba I and Xho I/Spe I, the light chain §Ügenes and heavy chain genes Fd were inserted into the phagemid vector pComb3Hª²SS successively and then electroporated into E.coli XL1ª²Blue. The specific Fab phage antibody library against Raji cell strain in human Bª²lymphoma 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¡Á107 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 Bª²lymphoma immunotherapy. ......