Identification of Human Lung Cancer-related Epitopes Using Phage display Peptide Library
10.3321/j.issn:1000-467X.2001.01.008
- VernacularTitle:应用噬菌体随机肽库研究人肺癌相关抗原表位
- Author:
Xin-Yuan LI
1
;
Ying-Wu HUANG
;
Yan-Hua ZHENG
;
Rui-Gang HAN
Author Information
1. 306 Hospital
- From:Chinese Journal of Cancer
2001;20(1):35-37
- CountryChina
- Language:Chinese
-
Abstract:
Objectives： The current study was designed to identify mimic polypeptide epitopes of lung cancer-related antigen WLA-Ag1 through screening random peptide library which was on display in phages, and provide useful information for further study of the characterization of WLA-Ag1 antigen and the potential chance for developing new diagnostic or therapeutic methods for lung cancer. Methods： Through affinity enrichment and immunoscreening of phage-displayed fifteen-peptide libraries with a monoclonal antibody named WLA2C4 against WLA-Ag1, the enriched phages was obtained and plated on agar plates. Some positive clones picked at random were confirmed by ELISA using WLA2C4 monoclonal antibody and DNA sequencing. The homologous comparison of amino acid sequences of positive clones was conducted through searching the protein sequence databank on Internet. The digestion analyses of amino acid sequences were also conducted by a software. Results： Twelve positive clones have been found with a consensual DNA sequence, that is, 5′ -GCT GGT TGG ATT ACT TTT CAT CGT CGT CAT CAT GAT CGT GTT CTT-3′ . Amino acid sequence was AGWITPHRRHHDRVL, with three trypsinase-digestible sites. This coincided with our previous finding that WLA-Ag1 can be digested by trypsinase. Conclusions： The amino acid sequence found in this study might be the epitope of WLA-Ag1. Screening of bioactive peptides from random peptide libraries using monoclonal antibodies as the ligands is an effective method to identify the epitopes of cancer-related antigens.
