Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA reference

The detection of antimitochondrial antibodies (AMA) is the specific diagnostic marker for primary biliary cholangitis. Indeed, it is the most specific autoantibody in clinical autoimmunity, with a high titer directed response to the inner lipoyl domain of PDC-E2. The current international reference...

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Main Authors: Zhuye Qin, Fangming Cheng, Mingming Zhang, Ruonan Qian, Hong Chen, Yaqin Zhao, Youtao Zhang, Yaping Dai, Chaochao Tang, Peng Jiang, Xiaoli Hua, Shen Li, Bing Zheng, Pin Yu, Xingjuan Shi, Suraj Timilsina, M. Eric Gershwin, Xiangdong Liu, Chungen Qian, Fang Qiu
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Journal of Translational Autoimmunity
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Online Access:http://www.sciencedirect.com/science/article/pii/S2589909024000327
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author Zhuye Qin
Fangming Cheng
Mingming Zhang
Ruonan Qian
Hong Chen
Yaqin Zhao
Youtao Zhang
Yaping Dai
Chaochao Tang
Peng Jiang
Xiaoli Hua
Shen Li
Bing Zheng
Pin Yu
Xingjuan Shi
Suraj Timilsina
M. Eric Gershwin
Xiangdong Liu
Chungen Qian
Fang Qiu
author_facet Zhuye Qin
Fangming Cheng
Mingming Zhang
Ruonan Qian
Hong Chen
Yaqin Zhao
Youtao Zhang
Yaping Dai
Chaochao Tang
Peng Jiang
Xiaoli Hua
Shen Li
Bing Zheng
Pin Yu
Xingjuan Shi
Suraj Timilsina
M. Eric Gershwin
Xiangdong Liu
Chungen Qian
Fang Qiu
author_sort Zhuye Qin
collection DOAJ
description The detection of antimitochondrial antibodies (AMA) is the specific diagnostic marker for primary biliary cholangitis. Indeed, it is the most specific autoantibody in clinical autoimmunity, with a high titer directed response to the inner lipoyl domain of PDC-E2. The current international reference for AMA detection is based upon sera samples of PBC patients. In rheumatic diseases, i.e. rheumatoid arthritis, great efforts are placed at development of international standards. In this study, we report the development of a monoclonal chimeric IgG1 antibody as a reference for AMA testing. A monoclonal 4G6 antibody was constructed from a murine monoclonal antibody specific for the inner lipoyl domain (ILD) of PDC-E2, by combining the variable region with the constant region of human IgG1. The 4G6 antibody recognizes all AMA epitopes containing the ILD of PDC-E2, including the classical BPO recombinant antigen in all currently available diagnostic methods. The binding affinity of the 4G6 antibody to PDC-E2 and BPO antigen reaches KD value of 7.22 × 10−11 M and 4.55 × 10−11 M, which is sufficient to use as a quantitative reference for all AMA tests. The unlimited availability of the 4G6 antibody makes it a promising candidate for use as an AMA reference or assay calibrator for the international community.
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spelling doaj-art-bed632776dd44a1a8d275f737fae3f5e2025-08-20T01:59:34ZengElsevierJournal of Translational Autoimmunity2589-90902024-12-01910026210.1016/j.jtauto.2024.100262Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA referenceZhuye Qin0Fangming Cheng1Mingming Zhang2Ruonan Qian3Hong Chen4Yaqin Zhao5Youtao Zhang6Yaping Dai7Chaochao Tang8Peng Jiang9Xiaoli Hua10Shen Li11Bing Zheng12Pin Yu13Xingjuan Shi14Suraj Timilsina15M. Eric Gershwin16Xiangdong Liu17Chungen Qian18Fang Qiu19Department of Laboratory Medicine, Southeast University Hospital, Nanjing, Jiangsu, 210096, ChinaReagent R&D Center, Shenzhen YHLO Biotech Co., Ltd, Shenzhen, Guangdong, 518116, ChinaKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, ChinaKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, ChinaKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, ChinaKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, ChinaKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, ChinaDepartment of Laboratory Medicine, The Fifth People's Hospital of Wuxi, Wuxi, Jiangsu, 214000, ChinaHangzhou Promise Biotech Inc., Hangzhou, Zhejiang, 310052, ChinaKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, ChinaDepartment of Laboratory Medicine, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210031, ChinaDepartment of Laboratory Medicine, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210031, ChinaDepartment of Laboratory Medicine, Shanghai Jiao Tong University School of Medicine Affiliated Renji Hospital, Shanghai, 200001, ChinaHangzhou Promise Biotech Inc., Hangzhou, Zhejiang, 310052, ChinaKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, ChinaDivision of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, Davis, CA, 95616, USADivision of Rheumatology, Allergy and Clinical Immunology, University of California at Davis School of Medicine, Davis, CA, 95616, USAKey Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, Nanjing, Jiangsu, 210088, China; Corresponding author. Key Laboratory of Developmental Genes and Human Diseases, School of Life Science and Technology, Southeast University, 2 Dongda Road, Nanjing, Jiangsu, 210088, China.Reagent R&D Center, Shenzhen YHLO Biotech Co., Ltd, Shenzhen, Guangdong, 518116, China; Corresponding author. Reagent R&D Center, Shenzhen YHLO Biotech Co., Ltd, Shenzhen, Guangdong, 518116, China.Department of Laboratory Medicine, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210031, China; Corresponding author. Department of Laboratory Medicine, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210031, China.The detection of antimitochondrial antibodies (AMA) is the specific diagnostic marker for primary biliary cholangitis. Indeed, it is the most specific autoantibody in clinical autoimmunity, with a high titer directed response to the inner lipoyl domain of PDC-E2. The current international reference for AMA detection is based upon sera samples of PBC patients. In rheumatic diseases, i.e. rheumatoid arthritis, great efforts are placed at development of international standards. In this study, we report the development of a monoclonal chimeric IgG1 antibody as a reference for AMA testing. A monoclonal 4G6 antibody was constructed from a murine monoclonal antibody specific for the inner lipoyl domain (ILD) of PDC-E2, by combining the variable region with the constant region of human IgG1. The 4G6 antibody recognizes all AMA epitopes containing the ILD of PDC-E2, including the classical BPO recombinant antigen in all currently available diagnostic methods. The binding affinity of the 4G6 antibody to PDC-E2 and BPO antigen reaches KD value of 7.22 × 10−11 M and 4.55 × 10−11 M, which is sufficient to use as a quantitative reference for all AMA tests. The unlimited availability of the 4G6 antibody makes it a promising candidate for use as an AMA reference or assay calibrator for the international community.http://www.sciencedirect.com/science/article/pii/S2589909024000327Primary biliary cholangitisAMAChimeric antibodyPDC-E2Inner lipoyl domain
spellingShingle Zhuye Qin
Fangming Cheng
Mingming Zhang
Ruonan Qian
Hong Chen
Yaqin Zhao
Youtao Zhang
Yaping Dai
Chaochao Tang
Peng Jiang
Xiaoli Hua
Shen Li
Bing Zheng
Pin Yu
Xingjuan Shi
Suraj Timilsina
M. Eric Gershwin
Xiangdong Liu
Chungen Qian
Fang Qiu
Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA reference
Journal of Translational Autoimmunity
Primary biliary cholangitis
AMA
Chimeric antibody
PDC-E2
Inner lipoyl domain
title Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA reference
title_full Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA reference
title_fullStr Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA reference
title_full_unstemmed Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA reference
title_short Development of a standardized monoclonal antibody to the inner lipoyl domain of PDC-E2 as a potential international AMA reference
title_sort development of a standardized monoclonal antibody to the inner lipoyl domain of pdc e2 as a potential international ama reference
topic Primary biliary cholangitis
AMA
Chimeric antibody
PDC-E2
Inner lipoyl domain
url http://www.sciencedirect.com/science/article/pii/S2589909024000327
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