caa a22 4500 605506221 CHVBK 20210128100626.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-015-6602-5 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6602-5 A monoclonal antibody-based VZV glycoprotein E quantitative assay and its application on antigen quantitation in VZV vaccine [Elektronische Daten] [Jian Liu, Rui Zhu, Xiangzhong Ye, Lianwei Yang, Yongmei Wang, Yanying Huang, Jun Wu, Wei Wang, Jianghui Ye, Yimin Li, Qinjian Zhao, Hua Zhu, Tong Cheng, Ningshao Xia] Varicella-zoster virus (VZV) is a highly infectious agent that causes varicella and herpes zoster (HZ), which may be associated with severe neuralgia. Vaccination is the most effective way to reduce the burden of the diseases. VZV glycoprotein E (gE) is the major and most immunogenic membrane protein that plays important roles in vaccine efficacy. A quantitative assay for gE content is desirable for the VZV vaccine process monitoring and product analysis. In this study, 70 monoclonal antibodies (mAbs) were obtained after immunizing mice with purified recombinant gE (rgE). The collection of mAbs was well-characterized, and a pair of high-affinity neutralization antibodies (capture mAb 4A2 and detection mAb 4H10) was selected to establish a specific and sensitive sandwich enzyme-linked immunosorbent assay (ELISA) to quantify the native and recombinant gE. The detection limit of this assay was found to be 1.95ng/mL. Furthermore, a reasonably good correlation between the gE content (as measured by the mAb-based quantitative ELISA) and the virus titer (as measured by the "gold standard” plaque assay) was observed when both assays were performed for tracking the kinetics of virus growth during cell culture. A total of 16 batches of lyophilized VZV vaccine were tested using the newly developed quantitative ELISA and classical plaque assay, demonstrating reasonably good correlation between gE content and virus titer. Therefore, this mAb-based gE quantitative assay serves as a rapid, stable, and sensitive method for monitoring viral antigen content, one additional quantitative method for VZV vaccine process and product characterization. This quantitative ELISA may also serve as a complementary method for virus titering. Springer-Verlag Berlin Heidelberg, 2015 Varicella-zoster virus (VZV) nationallicence VZV vaccine nationallicence Glycoprotein E nationallicence Sandwich ELISA nationallicence Virus titer nationallicence Liu Jian State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut Zhu Rui National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut Ye Xiangzhong Beijing Wantai Biological Pharmacy Enterprise, 102206, Beijing, China aut Yang Lianwei Beijing Wantai Biological Pharmacy Enterprise, 102206, Beijing, China aut Wang Yongmei State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut Huang Yanying National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut Wu Jun State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut Wang Wei State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut Ye Jianghui National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut Li Yimin Beijing Wantai Biological Pharmacy Enterprise, 102206, Beijing, China aut Zhao Qinjian National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut Zhu Hua Department of Microbiology and Molecular Genetics, New Jersey Medical School, Rutgers University, 225 Warren Street, 070101, Newark, NJ, USA aut Cheng Tong State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut Xia Ningshao State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4845-4853 0175-7598 99:11<4845 2015 99 253 https://doi.org/10.1007/s00253-015-6602-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6602-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liu Jian State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Zhu Rui National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Ye Xiangzhong Beijing Wantai Biological Pharmacy Enterprise, 102206, Beijing, China aut NATIONALLICENCE 700 1- Yang Lianwei Beijing Wantai Biological Pharmacy Enterprise, 102206, Beijing, China aut NATIONALLICENCE 700 1- Wang Yongmei State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Huang Yanying National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Wu Jun State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Wang Wei State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Ye Jianghui National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Li Yimin Beijing Wantai Biological Pharmacy Enterprise, 102206, Beijing, China aut NATIONALLICENCE 700 1- Zhao Qinjian National Institute of Diagnostics and Vaccine Development in infectious diseases, School of Public Health, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Zhu Hua Department of Microbiology and Molecular Genetics, New Jersey Medical School, Rutgers University, 225 Warren Street, 070101, Newark, NJ, USA aut NATIONALLICENCE 700 1- Cheng Tong State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 700 1- Xia Ningshao State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Life Sciences, Xiamen University, 361102, Xiamen, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/11(2015-06-01), 4845-4853 0175-7598 99:11<4845 2015 99 253