Aim: Preparation of monoclonal antibody (mAb) against GP73 protein.
Methods: The N-terminal peptide (AAAERGAVELK) of GP73 protein was displayed on T7 phage, the recombinant phage was amplified and used as the immunogen to immunize mouse to produce antibody. The titer of the antiserum and the positive hybridoma clones which secreted the mAb against GP73 protein were detected by ELISA. The mAb specificity was assayed by ELISA and Western blot.
Results: The high specificity mAb against GP73 protein was selected from the mouse immunized with the recombinant T7 phages displaying the epitope of GP73 by cell fusion and screening.
Conclusion: The appropriate protein epitope displayed on T7 phage could be used as alternative antigen to immunize animals to make specific antibody against the corresponding native protein.
Download full-text PDF |
Source |
---|
Anal Chim Acta
December 2018
Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian, 350116, China. Electronic address:
Golgi protein 73 (GP73) is a potential hepatocellular carcinoma serum marker with better sensitivity and specificity. In this study, a highly sensitive electrochemical immunosensor which combines the highly selective proximity ligation assay (PLA) and highly efficient enzyme-powered recycling amplification has been developed for GP73 determination. PLA is triggered by affinity binding of two labelled antibody-DNA (P1-RAb and P2-MAb) to target protein, resulting in increased specificity.
View Article and Find Full Text PDFAsian Pac J Cancer Prev
December 2015
Department of Laboratory Medicine, The Second Affiliated Hospital, Nanjing Medical University, Nanjing, China E-mail :
Background: Serum Golgi protein 73 (GP73) as a novel and potential marker for diagnosing hepatocellular carcinoma (HCC) have been found to be elevated in HCC patients and associated with clinical variables representing tumor growth and invasiveness. The aim of this study was to prepare a pair of monoclonal antibodys (mAbs) against GP73 and develop a newly designed double-antibody sandwich enzyme-linked immunosorbent assay (s-ELISA), which would be used in the detection of serum GP73 (sGP73) as well as in the diagnosis of HCC.
Materials And Methods: Produced by prokaryotic expression, the purified recombinant GP73 (rGP73), produced by prokaryotic expression, was used to immunize the Balb/c mice.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi
August 2013
Center of Clinical Laboratory, The No. 302 Hospital of The PLA, Beijing 100039, China.
Objective: To clone and express human Golgi glycoprotein73 protein, and prepare the monoclonal antibody (mAb) against the protein.
Methods: GP73 gene was amplified from HepG2 cells by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-GP73 and transformed into E. coli BL21.
J Biomed Res
November 2012
Department of Pharmacology, School of Pharmacy, Nanjing Medical University, Nanjing, Jiangsu 210029, China;
Recently, serum Golgi protein 73 (GP73) levels have been found to be elevated in patients with hepatocellular carcinoma (HCC), and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent assay (ELISA). Therefore, an anti-GP73 antibody with high specificity was highly demanded.
View Article and Find Full Text PDFXi Bao Yu Fen Zi Mian Yi Xue Za Zhi
July 2010
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing 102206, China.
Aim: Preparation of monoclonal antibody (mAb) against GP73 protein.
Methods: The N-terminal peptide (AAAERGAVELK) of GP73 protein was displayed on T7 phage, the recombinant phage was amplified and used as the immunogen to immunize mouse to produce antibody. The titer of the antiserum and the positive hybridoma clones which secreted the mAb against GP73 protein were detected by ELISA.
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!