Development and validation of a quantitative competitive ELISA for potency testing of equine anti rabies sera with other potential use |
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| Affiliation: | 1. Agence Nationale de Sécurité du Médicament et des Produits de Santé Laboratory Controls Division, Batch Release and Marketing Surveillance of Biological Products Department, 321 Avenue Jean Jaurès, F-69007 Lyon, France;2. Biological Controls of Immunological Products and Biological Security Department, 321 Avenue Jean Jaurès, F-69007 Lyon, France;3. Antiviral Strategies Unit, Virology Department, Institut Pasteur, 25, rue du Docteur Roux, F-75724 Paris Cedex 15, France;1. Viral Pseudotype Unit (Medway), School of Pharmacy, University of Kent, Chatham Maritime, Kent, UK;2. Viral Pseudotype Unit (Fitzrovia), Faculty of Science and Technology, University of Westminster, London, UK;1. West China Hospital Emergency Department (WCHED), State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Collaborative Innovation Center of Biotherapy, Chengdu, Sichuan 610041, China;2. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;3. Division of Arboviral Vaccines, National Institutes of Food and Drug Control (NIFDC), Beijing 100050, China;4. Disaster Medicine Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China;5. Savaid Medical School, University of Chinese Academy of Sciences, Beijing 100049, China;6. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (China CDC), Beijing 102206, China;7. Research Network of Immunity and Health (RNIH), Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing 100101, China;1. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian 361005, PR China;2. School of Public Health, Xiamen University, Xiamen, Fujian 361005, PR China;3. School of Life Science, Xiamen University, Xiamen, Fujian 361005, PR China;4. Xiamen Innovax Biotech Co., Ltd, Xiamen, Fujian 361005, PR China |
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| Abstract: | In case of a bite by a rabies infected animal, the World Health Organisation recommends a prophylactic treatment including the administration of Human Rabies Immunoglobulins (HRIGs) or highly purified F(ab′)2 fragments produced from Equine Rabies Immunoglobulin (F(ab′)2 – ERIGs). According to international regulation, quality control of F(ab′)2 – ERIGs lots requires potency testing by the in vivo Mouse Neutralisation Test (MNT) prior marketing. However, the strategy of the 3Rs (Reduce, Refine, Replace) for animal testing required by the European Directive encourages the replacement of the in vivo potency test by an in vitro assay. In this context, a competitive ELISA method (c-ELISA) has been developed by the Agence Nationale de Sécurité du Médicament et des Produits de Santé where F(ab′)2 – ERIGs are in competition with a monoclonal antibody recognizing the trimeric native form of the rabies glycoprotein. After a full validation study, the c-ELISA has been applied to commercial batches of F(ab′)2 – ERIGs. A correlation study with the MNT demonstrated a similarity between the two methods (r = 0.751). Moreover, the c-ELISA method which does not need any species specific reagent has been applied to HRIGs potency testing as an alternative method to Rapid Fluorescent Focus Inhibition Test (RFFIT), thus avoiding the handling of live rabies virus in BSL3 containment. In conclusion, the c-ELISA has shown its potential to replace MNT and possibly RFFIT for the quantification of rabies immunoglobulin. After optimisation it may be used for the quantification of rabies immunoglobulin in any animal species, notably for rabies immunogenicity assay in mice. |
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| Keywords: | Competitive ELISA Rabies immunoglobulins (ERIGs, HRIGs) Quality control Validation 3Rs Batch release |
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