Original Research
Copyright ©The Author(s) 2001. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Dec 15, 2001; 7(6): 811-815
Published online Dec 15, 2001. doi: 10.3748/wjg.v7.i6.811
Construction and selection of the natural immune Fab antibody phage display library from patients with colorectal cancer
Bao-Ping Wu, Bing Xiao, Tian-Mo Wan, Ya-Li Zhang, Zhen-Shu Zhang, Dian-Yuan Zhou, Zhuo-Sheng Lai, Chun-Fang Gao
Bao-Ping Wu, Bing Xiao, Tian-Mo Wan, Ya-Li Zhang, Zhen-Shu Zhang, Dian-Yuan Zhou, Zhuo-Sheng Lai, Institute for Digestive Diseases, Nanfang Hospital, Guangzhou 510515, Guangdong Province, China
Chun-Fang Gao, Surgical Department of Colon and Rectum, 150 Central Hospital, Luoyang 471031, Henan Province, China
Author contributions: All authors contributed equally to the work.
Supported by the Natural Science Foundation of Guangdong Provin ce, China, No.980120 and the Foundation of Excellent Youth Teacher, China, 2001
Correspondence to: Dr. Bao-Ping Wu, Institute for Digestive Medicine, Nanfang Hospital, Guangzhou 510515, Guangdong Province, China. bpwu@263.net
Telephone: +86-20-85141544
Received: April 6, 2001
Revised: June 1, 2001
Accepted: June 15, 2001
Published online: December 15, 2001
Abstract

AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer.

METHODS: Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer. RT-PCR was used to amplify the heavy chain Fd and light chain κ and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected.

RESULTS: The amplified fragments of Fd and κ gained by RT-PCR were about 650 bp. Fd and κ PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 × 106. The libraries were enriched about 120-fold by 3 cycles of adsorption-elution-multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5 clones of Fab phage anti bodies which had binding activities with antigens of colorectal cancer.

CONCLUSION: The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer.

Keywords: colorectal neoplasms; immunology; bacterio phages; antibody library