Cloning, expression, purification, and characterization of LC-1 ScFv with GFP tag

• Published in: Journal of Zhejiang University. B. Science Vol. 6; no. 8; pp. 832 - 837
• Main Author: 卢敏 龚兴国 于红 郦剑勇
• Format: Journal Article
• Language: English
• Published: China Institute of Biologic Macromolecule and Enzyme Engineering, School of Life Science, Zhejiang University, Hangzhou 310027, China 01.08.2005; Zhejiang University Press
• Subjects: Adenocarcinoma - immunology; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - isolation & purification; Cell Line; Cell Line, Tumor; Cloning, Molecular - methods; GFP; Green Fluorescent Proteins - biosynthesis; Green Fluorescent Proteins - genetics; Humans; Hybridomas - metabolism; Immunoglobulin Fragments - biosynthesis; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - immunology; Immunoglobulin Fragments - isolation & purification; Lung Neoplasms - immunology; Plant & Animal Sciences and Biotechnology; Protein Engineering - methods; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - isolation & purification; RNA; 克隆技术; 基因工程; 绿色荧光蛋白质; 遗传表达; Protein fusion; ScFv (single chain variable fragment); Purification; GFP (green fluorescent protein) tag

Total RNA was isolated from the hybridoma cell line （LC- 1 ）, which secretes anti-lung adenocarcinoma monoclonal antibody, and was transferred into cDNA. Based on the FRl （framework region l） and FR4 conserved regions of LC-1 gene, the variable regions of heavy chain （Vh） and light chain （Vl） were a...