Increased sensitivity of gp210 autoantibody detection using a newly designed gp210 antigen

Jing Xu; Haoyi Zhang; Chan Wang; Peng Jiang; Chongxu Han; Yaping Dai; Fang Qiu; Yuhua Gong; Yuzhang Jiang; Ping Xu; Mingming Zhang; Luyao Zhang; Xingjuan Shi; Sufang Chen; Ye Tian; Michael F Seldin; M Eric Gershwin; Xiangdong Liu; Li Li

doi:10.1016/j.jim.2021.113211

Increased sensitivity of gp210 autoantibody detection using a newly designed gp210 antigen

J Immunol Methods. 2022 Feb:501:113211. doi: 10.1016/j.jim.2021.113211. Epub 2021 Dec 28.

Authors

Jing Xu¹, Haoyi Zhang², Chan Wang³, Peng Jiang², Chongxu Han⁴, Yaping Dai⁵, Fang Qiu⁶, Yuhua Gong⁷, Yuzhang Jiang⁸, Ping Xu⁹, Mingming Zhang², Luyao Zhang², Xingjuan Shi², Sufang Chen⁹, Ye Tian¹⁰, Michael F Seldin¹¹, M Eric Gershwin¹², Xiangdong Liu¹³, Li Li¹⁴

Affiliations

¹ Institute of Life Sciences and Technology, Southeast University, 2 Sipailou Road, Nanjing, Jiangsu 210096, China; Department of Clinical Laboratory, ZhongDa Hospital, Southeast University, Nanjing, Jiangsu 210009, China.
² Institute of Life Sciences and Technology, Southeast University, 2 Sipailou Road, Nanjing, Jiangsu 210096, China.
³ Institute of Translational Medicine, Medical College, Yangzhou University, 136 Yangjiang Middle Road, Yangzhou, Jiangsu 225001, China.
⁴ Department of Laboratory Medicine, Subei People's Hospital, Clinical Medical College, Yangzhou University, 98 Nantong West Road, Yangzhou, Jiangsu 225001, China.
⁵ Department of Laboratory Medicine, The Fifth People's Hospital of Wuxi, 1215 Guangrui Road, Wuxi, Jiangsu 214000, China.
⁶ Department of Laboratory Medicine, The Forth Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210031, China.
⁷ Department of Laboratory Medicine, The Third People's Hospital of Zhenjiang, 300 Daijiamen, Zhenjiang, Jiangsu 212021, China.
⁸ Department of Laboratory Medicine, Huai'an First People's Hospital, Nanjing Medical University, 1 Huanghe West Road, Huai'an, Jiangsu 223300, China.
⁹ Department of Laboratory Medicine, The Fifth People's Hospital of Suzhou, Soochow University, 10 Guangqian Road, Suzhou, Jiangsu 215131, China.
¹⁰ Department of Radiology, The Second Affiliated Hospital of Soochow University，1055 Sanxiang Road, Suzhou, Jiangsu 215004, China.
¹¹ Department of Biochemistry and Molecular Medicine, University of California at Davis School of Medicine, 4453 Tupper Hall, Davis, CA 95616, USA; Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, School of Medicine, Genome and Biomedical Sciences Facility Building, 451 Health Sciences Drive, Suite 6510, Davis, CA 95616, USA.
¹² Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, School of Medicine, Genome and Biomedical Sciences Facility Building, 451 Health Sciences Drive, Suite 6510, Davis, CA 95616, USA.
¹³ Institute of Life Sciences and Technology, Southeast University, 2 Sipailou Road, Nanjing, Jiangsu 210096, China. Electronic address: xiangdongliu@seu.edu.cn.
¹⁴ Institute of Life Sciences and Technology, Southeast University, 2 Sipailou Road, Nanjing, Jiangsu 210096, China; Department of Clinical Laboratory, ZhongDa Hospital, Southeast University, Nanjing, Jiangsu 210009, China. Electronic address: lilee_immuno@seu.edu.cn.

PMID: 34971632
DOI: 10.1016/j.jim.2021.113211

Abstract

Objectives: The detection of autoantibody to glycoprotein 210 (gp210 Ab) against a 15 amino-acid peptide epitope by enzyme-linked immunosorbent assay (ELISA) has been widely used in the diagnosis of primary biliary cholangitis (PBC). However, this small peptide antigen presents spatial limitations for antibody access, which reduces the sensitivity of autoantibody detection. A recombinant gp210 antigen was constructed for increased sensitivity in antibody detection is described here.

Methods: The gp210 C terminal 18 amino acid coding sequence was ligated to the modified C-terminal 108 amino acid coding sequence of human serum albumin (mHSA108) and produced as a recombinant gp210 antigen mHSA108-gp210-C18. Measurements of gp210 Ab using the gp210 C-terminal 25 amino acid peptide (gp210-C25) and mHSA108-gp210-C18 by in-house ELISA were compared. ELISAs with mHSA108-gp210-C18 and commercial INOVA kit for gp210 Ab detection were also compared in PBC patients and healthy controls. The correlation between the two assays was analyzed and their efficiency in diagnosing was compared.

Results: Of 86 PBC samples, 35 (40.70%) and 44 (52.33%) positive samples were detected for anti-gp210 Ab using gp210-C25 and mHSA108-gp210-C18, respectively. Of 252 samples from PBC, 114 (45.24%) were positive for mHSA108-gp210-C18 ELISA whereas 94 (37.3%) for commercial ELISA (INOVA). All positive samples detected with commercial ELISA kit were also tested positive in mHSA108-gp210-C18 ELISA. Among 374 patients with other autoimmune diseases, anti-gp210 Ab were detected by mHSA108-gp210-C18 ELISA in 0.95% systemic lupus erythematosus (SLE) patients (2/210), 13.04% rheumatoid arthritis (RA) patients (13/97), and 1.47% of Sjögren's Syndrome (SS) patients (1/67).

Conclusions: Compared to the gp210 peptide antigen, the sensitivity of the ELISA system using mHSA108-gp210-C18 antigen was improved. The novel gp210 antigen could be useful for screening patients known to be at increased risk of developing PBC.

Keywords: Enzyme-linked immunosorbent assay; Glycoprotein 210; Human serum albumin; Primary biliary cholangitis.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Adult
Aged
Aged, 80 and over
Autoantibodies / blood*
Autoantigens / immunology*
Biomarkers / blood
Case-Control Studies
Enzyme-Linked Immunosorbent Assay*
Epitopes
Female
Humans
Liver Cirrhosis, Biliary / blood
Liver Cirrhosis, Biliary / diagnosis*
Liver Cirrhosis, Biliary / immunology
Male
Middle Aged
Nuclear Pore Complex Proteins / immunology*
Peptide Fragments / immunology*
Predictive Value of Tests
Reproducibility of Results
Young Adult

Substances

Autoantibodies
Autoantigens
Biomarkers
Epitopes
NUP210 protein, human
Nuclear Pore Complex Proteins
Peptide Fragments