Volume 17, No 7, Jul 2007

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 17 Issue 7, July 2007: 650-660

ORIGINAL ARTICLES

Generation and selection of immunized Fab phage display library against human B cell lymphoma

Yongmei Shen^1,2, Xiaochun Yang¹, Ningzheng Dong³, Xiaofang Xie^1,2, Xia Bai³ and Yizhen Shi^1,2

¹Department of Radioimmunoassay, Second Affiliated Hospital of Soochow University, Suzhou 215004, China

²Department of Clinical Laboratory, Second Affiliated Hospital of Soochow University, Suzhou 215004, China

³Leukemia Research Unit, Jiangsu Institution of Hematology, First Affiliated Hospital of Soochow University, Suzhou 215006, China Correspondence: Yongmei Shen(yongmei_shen@yahoo.com)

The approval of using monoclonal antibodies as a targeted therapy in the management of patients with B cell lymphoma has led to new treatment options for this group of patients. Production of monoclonal antibodies by the traditional hybridoma technology is costly, and the resulting murine antibodies often have the disadvantage of triggering human anti-mouse antibody (HAMA) response. Therefore recombinant Fab antibodies generated by the phage display technology can be a suitable alternative in managing B cell lymphoma. In this study, we extracted total RNA from spleen cells of BALB/c mice immunized with human B lymphoma cells, and used RT-PCR to amplify cDNAs coding for the κ light chains and Fd fragments of heavy chains. After appropriate restriction digests, these cDNA fragments were successively inserted into the phagemid vector pComb3H-SS to construct an immunized Fab phage display library. The diversity of the constructed library was approximately 1.94×10⁷. Following five rounds of biopanning, soluble Fab antibodies were produced from positive clones identified by ELISA. From eight positive clones, FabC06, FabC21, FabC43 and FabC59 were selected for sequence analysis. At the level of amino acid sequences, the variable heavy domains (V_H) and variable light domains (V_L) were found to share 88-92% and 89-94% homology with sequences coded by the corresponding murine germline genes respectively. Furthermore, reactivity with membrane proteins of the B cell lymphoma was demonstrated by immunohistochemistry and western blotting. These immunized Fab antibodies may provide a valuable tool for further study of B cell lymphoma and could also contribute to the improvement of disease therapy

Cell Research (2007) 17:650-660. doi: 10.1038/cr.2007.57; published online 10 July 2007

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