Gastric Cancer
Copyright ©The Author(s) 2002. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Aug 15, 2002; 8(4): 619-623
Published online Aug 15, 2002. doi: 10.3748/wjg.v8.i4.619
Identification of tumor associated single-chain Fv by panning and screening antibody phage library using tumor cells
Yong-Zhan Nie, Feng-Tian He, Zhi-Kui Li, Kai-Chun Wu, Yun-Xin Cao, Bao-Jun Chen, Dai-Ming Fan
Yong-Zhan Nie, Feng-Tian He, Zhi-Kui Li, Kai-Chun Wu, Bao-Jun Chen, Dai-Ming Fan, Yun -Xin Cao, Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China
Author contributions: All authors contributed equally to the work.
Supported by the National “63” Strategy for Science (102-10-01-06) and National Natural Science Foundation of China, No.39525020
Correspondence to: Dai-Ming Fan, Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, Changle West Road, Xi'an 710032, Shannxi Province China. fandaim@fmmu.edu.cn
Telephone: +86-29-2539041 Fax: +86-29-2539041
Received: June 12, 2001
Revised: September 2, 2001
Accepted: September 14, 2001
Published online: August 15, 2002
Abstract

AIM: To study the feasibility of panning and screening phage-displaying recombinant single-chain variable fragment (ScFv) of anti-tumor monoclonal antibodies for fixed whole cells as the carriers of mAb-binding antigens.

METHODS: The recombinant phage displaying libraries for anti-colorectal tumor mAb MC3Ab, MC5Ab and anti-gastric tumor mAb MGD1 was constructed. Panning and screening were carried out by means of modified fixation of colorectal and gastric tumor cells expressed the mAb-binding antigens. Concordance of binding specificity to tumor cells between phage clones and parent antibodies was analyzed. The phage of positive clones was identified with competitive ELISA, and infected by E. coli HB2151 to express soluble ScFv.

RESULTS: The ratio of positive clones to MC3-ScF-MC5-ScFv and MGD1-ScFv were 60%, 24% and 30%. MC3-ScFv had Mr 32000 confirmed by Western blot. The specificity to antigen had no difference between 4 positive recombinant phage antibodies and MC3Ab.

CONCLUSION: The modified process of fixing whole tumor cells is efficient, convenient and feasible to pan and screen the phage-displaying ScFv of anti-tumor monoclonal antibodies.

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