Construction and screening of natural immune Fab antibody phage library from patients with colorectal cancer
WU Bao-ping
ZHANG Ya-li
XIAO Bing
HANG Zhen-shu
LAI Zhuo-Sheng
WANG Ya-dong
GAO Chun-Fang
ZHOU Dian-yuan
Abstract：Objective To construct natural immune Fab fragment phage display library and to screen the antibodies against human colorectal cancer. Methods Total RNA in the metastatic lymph nodes from patients with colorectal cancer was extracted routinely, and reverse transcriptase PCR employed to amplify the heavy chain Fd and light chain κ cDNA. The amplification products were inserted in succession into the vector pComb3 to construct human Fab fragment library, which was screened and identified using phage display technique. Results The heavy chain Fd fragment and light chain κ fragment (650 bp in length) were amplified, and the products were inserted into pComb3 vector with a recombination rate of 40%. The constructed phage display Fab library had a capacity of up to 1.48×106, and was enriched approximately 120 times after 3 cycles of panning. Dot immunoblotting found the expression of Fab fragment which could bind to the antigens of human colorectal cancer. Conclusion The phage display library of the antibody Fab fragment from naturally immunized lymph nodes is successfully established, rendering it possible to screen the antibodies against human colorectal cancer.
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Publication Date：2001-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 270-274 )