生物制品质量控制中动物试验的替换或移除指南

Adoptedon the recommendation of the Eighty-first meeting of the World HealthOrganization Expert Committee onBiological Standardization, 13–16October2025. Thisfinalized andediteddocumentis beingprovided here prior to its formalpublication both © World Health Organization 2025 All rights reserved. The designations employed and the presentation of the material in this document do not imply the expression ofany opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, The mention of specific companies or of certain manufacturers’ products does not imply that they are endorsedor recommended by the World Health Organization in preference to others of a similar nature that are not All reasonable precautions have been taken by the World Health Organization to verify theinformation containedin this document. However, the material is being distributed without warranty of any kind, either expressed orimplied. The responsibility for the interpretation and use of the material lies with the reader. In no event shall the Annex 2 Guidelines on thereplacement orremoval of animal tests for the qualitycontrol of biological products 1.Introduction 2.Purpose and scope 3.Terminology 6 4.General considerations 5.Adventitious agent testing105.1In vivo adventitious agent testing10 6.Pyrogenicity and endotoxin testing6.1Pyrogenicitytesting6.2Endotoxin testing 7.Neurovirulence testing7.1Yellow fever vaccine7.2Oral poliomyelitis vaccine 8.Potency testing8.1Conclusions 2023 9.Specific toxicity249.1Diphtheria, tetanus and acellular pertussis vaccines249.2Polysaccharide vaccines conjugated to a diphtheria or tetanus toxoid 10.Innocuity testing10.1Conclusions 11.Development and use of international and otherbiologicalreferencestandards 12.Guidance for national regulatory authorities Authors and acknowledgements References Appendix1 List of WHO documents for biological products publishedpriorto2018in which any mentionof the innocuity test(also knownasthe Important note Guidelines published by the World Health Organization (WHO) are intended to be scientificand advisory in nature. Each of the following sections constitutes guidance for nationalregulatory authorities (NRAs) and for manufacturers of biological products.If an NRA sodesires, theguidance given in theseWHO Guidelines may be adopted as definitive national The implementation of existing in vitro alternativesto animal-based testingfor thequality control of biological productsandthedevelopment of new in vitro methods,particularlywheregaps still remain,are strongly recommended.Towards this goal,internationalcooperationbetweenrelevant stakeholders,along with enhancedregulatory convergence,will Theguidanceprovided in the current document with regard to the use of in vitro tests,and by extension the replacement or removal of animal tests,isscience-based.Such guidanceshould beviewed as supersedingthecorresponding quality controlrecommendationsspecifiedin WHORecommendations, Guidelines and other guidancedocumentson biological productspublished prior to 2025. Product developers and manufacturers, and other stakeholders should Abbreviations CHOChinese hamster ovaryCNScentral nervous systemCQAcritical quality attributeELISAenzyme-linked immunosorbent assayEPOErythropoietinFNVFrench neurotropic vaccineGMPgood manufacturing practicesHTShigh-throughput sequencingICHInternational Council for Harmonisation of Technical Requirements forPharmaceuticals for HumanUseLALLimulusamoebocyte lysateMAPRECmutant analysis by PCR and restriction enzyme cleavage 1.Introduction Animal testing has longbeen animportanttoolinthe development ofmedicinalbiologicalproducts, providing critical information on their mechanisms of action, safety and efficacy. Insome cases,such testingalsocontinuesto be used post-approval to monitor product qualityaspart of the quality control processesofmanufacturers and national control laboratories(NCLs).However,thegrowingrecognition thatappropriate in vitrotestsarebetter suited forroutine Historically, efforts toconducthigh-quality scientific researchin the most humane wayhave been underpinned by the guiding principles of the“3Rs” approach of replacement,reduction and refinement(1). This approach aims to promote good practices and the humanetreatment of animals used in scientific research and product testing,including throughreductionof the number of animals used and refinement ofanimal test methodologies.However, while reduction and refinementeffortsmay helpto promoteanimal welfare, suchapproaches (in particular, reduction) can increasethevariability oftestresults.In contrast, thereplacement of animal tests with appropriate in vitro testseliminatesany requirement to useanimals while maintaining or improving the scientific relevance of thedataobtained.Theuse For more than 70 years, WHORecommendations, Guidelines and other guidancedocuments haveset the norms and standards forthe production, quality contr