Bibliography to support the adoption of 3Rs approaches in biologics quality control testing

The five sections below reflect the broad testing categories most commonly identified during the review of WHO testing guidelines. We have also included a ‘General’ section that provides the wider context for the 3Rs in quality control and batch release testing. 

• Publications under each category can be accessed by clicking on the category name in the list of page contents above.
• To return to the top of the page, use the up arrow in the grey circle in the bottom right corner of the page.
• To search the bibliography more specifically, for example by product type (e.g. poliovirus) or specific test/assay (e.g. monocyte activation test), use the ‘Ctrl+F’ keyboard shortcut.

Hoefnagel M et al, (2023). Rational arguments for regulatory acceptance of consistency testing: benefits of non-animal testing over in vivo release testing of vaccines. Expert Review of Vaccines 22(1): 369–377. doi: 10.1080/14760584.2023.2198601

Lilley E et al. (2023). Integrating 3Rs approaches in WHO guidelines for the batch release testing of biologicals: Responses from a survey of National Control Laboratories and National Regulatory Authorities. Biologicals 84: 101721 10.1016/j.biologicals.2023.101721

Dierick J-F et al, (2022). The consistency approach for the substitution of in vivo testing for the quality control of established vaccines: practical considerations and progressive vision. Open Res Europe 2: 116. doi: 10.12688/openreseurope.15077.2

Lilley E et al. (2022). Integrating 3Rs approaches in WHO guidelines for the batch release testing of biologicals: Responses from a survey of vaccines and biological therapeutics manufacturers. Biologicals 81: 101660 10.1016/j.biologicals.2022.11.002

Saint-Raymond A et al, (2022). Reliance is key to effective access and oversight of medical products in case of public health emergencies. Expert Rev Clin Pharmacol 15(7): 805-810. doi: 10.1080/17512433.2022.2088503

Lilley E et al. (2021). Integrating 3Rs approaches in WHO guidelines for the batch release testing of biologicals. Biologicals 74: 24-27 10.1016/j.biologicals.2021.10.002

Akkermans A et al, (2020). Animal testing for vaccines. Implementing replacement, reduction and refinement: challenges and priorities. Biologicals 68: 92-107. doi: 10.1016/j.biologicals.2020.07.010

Cuff P and Woods A (2019). Regulating Medicines in a Globalized World: The Need for Increased Reliance Among Regulators. National Academies Press (US). doi: 10.17226/25594.2020.  

Uhlrich S et al, (2018). 3Rs in Quality Control of Human Vaccines: Opportunities and Barriers. In: Kojima, H., Seidle, T., Spielmann, H. (eds) Alternatives to Animal Testing. Springer, Singapore. doi: 10.1007/978-981-13-2447-5_10

Bruysters M et al, (2017). Drivers and barriers in the consistency approach for vaccine batch release testing: Report of an international workshop. Biologicals 48: Pages 1-5. doi: 10.1016/j.biologicals.2017.06.006

Schutte K et al, (2017). Modern science for better quality control of medicinal products "Towards global harmonization of 3Rs in biologicals": The report of an EPAA workshop. Biologicals  48: Pages 55-65. doi: 10.1016/j.biologicals.2017.05.006 

Hendriksen C et al, (2008). The consistency approach for the quality control of vaccines. Biologicals 36(1): 73-7. doi: 10.1016/j.biologicals.2007.05.002

Halder M (2001). Three Rs potential in the development and quality control of immunobiologicals. Altex 18 Suppl 1: 13-47. PMID: 11854853

Thurman T et al, (2023). Comparison of pyrogen assays by testing products exhibiting low endotoxin recovery. ALTEX 40(1):117-124. doi: 10.14573/altex.2202021

Charton E (2022). European Pharmacopoeia Approach to Testing for Pyrogenicity. American Pharmaceutical Review July/August 2022. Link to article: https://www.americanpharmaceuticalreview.com

Etna M et al, (2022). Optimization of the monocyte activation test for evaluating pyrogenicity of tick-borne encephalitis virus vaccine. ALTEX 37(4): 532-544. doi: 10.14573/altex.2002252

Tindall B et al, (2021). Recombinant bacterial endotoxin testing: a proven solution. Bio Techniques 70(5): 290-300. doi: 10.2144/btn-2020-0165

Hartung T (2021). Pyrogen testing revisited on occasion of the 25th anniversary of the whole blood monocyte activation test. ALTEX 38(1): 3-19. doi: 10.14573/altex.2101051 

Bolden et al, (2020). Currently Available Recombinant Alternatives to Horseshoe Crab Blood Lysates: Are They Comparable for the Detection of Environmental Bacterial Endotoxins? A Review. PDA J Pharm Sci Technol 274(5): 602-611. doi: 10.5731/pdajpst.2020.012187

Gorman R (2020). Atlantic Horseshoe Crabs and Endotoxin Testing: Perspectives on Alternatives, sustainable Methods, and the 3Rs (Replacement, Reduction, and Refinement). Front Mar Sci 30(7): 1-11. doi: 10.3389%2Ffmars.2020.582132

Piehler et al, (2020). Comparison of LAL and rFC Assays-Participation in a Proficiency Test Program between 2014 and 2019. Microorganisms 8(3):418 doi: 10.3390/microorganisms8030418 

Vipond C et al, (2019). Development and validation of a monocyte activation test for the control/safety testing of an OMV-based meningococcal B vaccine. Vaccine 37(29): 3747-3753. dio: 10.1016/j.vaccine.2018.06.038

Kirfalusi-Gannon J et al, (2018). The Role of Horseshoe Crabs in the Biomedical Industry and Recent Trends Impacting Species Sustainability. Front Mar Sci 05: 1-13. dio: 10.3389/fmars.2018.00185

Bolden J and Smith K (2017). Application of Recombinant Factor C Reagent for the Detection of Bacterial Endotoxins in Pharmaceutical Products. PDA J Pharm Sci Technol 71(5): 405-412 dio: 10.5731/pdajpst.2017.007849

Fennrich S et al, (2016). More than 70 years of pyrogen detection: Current state and future perspectives. Altern Lab Anim 44(3): 239-53. dio: 10.1177/026119291604400305

Schindler S et al, (2009). Development, validation and applications of the monocyte activation test for pyrogens based on human whole blood. ALTEX 26(4): 265-277. dio: 10.14573/altex.2009.4.265

Ding J and Ho B (2001). A new era in pyrogen testing. Trends Biotechnol 19(8): 277-81. dio: 10.1016/s0167-7799(01)01694-8

Konz J et al, (2021). Evaluation and validation of next-generation sequencing to support lot release for a novel type 2 oral poliovirus vaccine. Vaccine X 8: 100102 doi: 10.1016/j.jvacx.2021.100102

May Fulton C and Bailey W (2021). Live Viral Vaccine Neurovirulence Screening: Current and Future Models. Vaccines (Basel) 9(7):710. doi: 10.3390/vaccines9070710

Charlton B et al, (2020). The Use of Next-Generation Sequencing for the Quality Control of Live-Attenuated Polio Vaccines. J Infect Dis 222(11):1920-1927. doi: 10.1093/infdis/jiaa299

da Costa A et al, (2018). Innovative in cellulo method as an alternative to in vivo neurovirulence test for the characterization and quality control of human live Yellow Fever virus. Biologicals 53:19-29. doi: 10.1016/j.biologicals.2018.03.004

Wood D (1999). Neurovirulence. Dev Biol Stand 101:127-9. PMID: 10566785

Horie H et al, (1998). Estimation of the neurovirulence of poliovirus by non-radioisotope molecular analysis to quantify genomic changes. Biologicals 26(4): 289-97. doi: 10.1006/biol.1998.0159

Chumakov K et al, (1993). Assessment of the viral RNA sequence heterogeneity for control of OPV neurovirulence. Dev Biol Stand 78:79-89. PMID: 8388834

Barone P et al, (2023). Historical evaluation of the in vivo adventitious virus test and its potential for replacement with next generation sequencing (NGS). Biologicals 81:101661. doi: 10.1016/j.biologicals.2022.11.003

Morris C et al, (2021). Adventitious agent detection methods in bio-pharmaceutical applications with a focus on viruses, bacteria, and mycoplasma. Curr Opin Biotechnol 71:105-114. doi: 10.1016/j.copbio.2021.06.027

Charlebois R et al, (2020). Sensitivity and breadth of detection of high-throughput sequencing for adventitious virus detection. npj Vaccines 5: 61. doi: 10.1038/s41541-020-0207-4

Khan A et al, (2020). Report of the second international conference on next generation sequencing for adventitious virus detection in biologics for humans and animals. Biologicals 67: 94-111. doi: 10.1016/j.biologicals.2020.06.002

Pennings J et al, (2020). A next-generation sequencing based method for determining genetic stability in Clostridium tetani vaccine strains. Biologicals 64: 10-14. doi: 10.1016/j.biologicals.2020.02.001

Gombold J et al, (2014). Systematic evaluation of in vitro and in vivo adventitious virus assays for the detection of viral contamination of cell banks and biological products. Vaccine 32(24): 2916-26. doi: 10.1016/j.vaccine.2014.02.021

Iwaki M et al, (2023). An ELISA system for tetanus toxoid potency tests: An alternative to lethal challenge. Biologicals 82:101681 doi: 10.1016/j.biologicals.2023.101681

Tedcastle R et al, (2022). Report on the WHO collaborative study to establish Universal Reagents for the D-Antigen potency testing of Inactivated Polio Vaccines. Link to article: https://www.who.int/publications/m/item/who-bs-2022.2432

Vandebriel R et al, (2022). Development of a cell line-based in vitro assay for assessment of Diphtheria, Tetanus and acellular Pertussis (DTaP)-induced inflammasome activation. Vaccine 40(38): 5601-5607. doi: 10.1016/j.vaccine.2022.08.022

Riches-Duit R et al, (2021). Characterisation of diphtheria monoclonal antibodies as a first step towards the development of an in vitro vaccine potency immunoassay. Biologicals. 69: 38-48. doi: 10.1016/j.biologicals.2020.12.002

Riches-Duit R et al, (2021). Characterisation of tetanus monoclonal antibodies as a first step towards the development of an in vitro vaccine potency immunoassay. Biologicals. 71: 31-41. doi: 10.1016/j.biologicals.2021.04.002

Signorazzi A et al, (2021). In vitro assessment of tick-borne encephalitis vaccine: Suitable human cell platforms and potential biomarkers. ALTEX 38(3): 431-441. doi: 10.14573/altex.2010081

Stalpers C et al, (2021). Variability of in vivo potency tests of Diphtheria, Tetanus and acellular Pertussis (DTaP) vaccines. Vaccine. 39(18): 2506-2516. doi: 10.1016/j.vaccine.2021.03.078

Ticha O et al, (2021). A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen. NPJ Vaccines 6: 88. 10.1038%2Fs41541-021-00344-1

Imura A et al, (2020). Development of an Enterovirus 71 Vaccine Efficacy Test Using Human Scavenger Receptor B2 Transgenic Mice. J Virol 94(6):e01921-19. doi: 10.1128/jvi.01921-19

Kouiavskaia D et al, (2020). Universal ELISA for quantification of D-antigen in inactivated poliovirus vaccines. Journal of Virological Methods 276: 113785. doi: 10.1016/j.jviromet.2019.113785

Michiels T et al, (2020). Degradomics-Based Analysis of Tetanus Toxoids as a Quality Control Assay. Vaccines  8(4): 712. doi: 10.3390/vaccines8040712

Rajam G et al, (2019). Development and validation of a robust multiplex serological assay to quantify antibodies specific to pertussis antigens. Biologicals 57: 9-20. doi: 10.1016/j.biologicals.2018.11.001

Morgeaux S et al, (2018). Replacement of in vivo human rabies vaccine potency testing by in vitro glycoprotein quantification using ELISA - Results of an international collaborative study. Vaccine 35(6): 966-971. doi: 10.1016/j.vaccine.2016.12.039

Chabaud-Riou M et al, (2017). G-protein based ELISA as a potency test for rabies vaccines. Biologicals 46: 124-129. doi: 10.1016/j.biologicals.2017.02.002

Byung-Chul K et al, (2016). A collaborative study of an alternative in vitro potency assay for the Japanese encephalitis vaccine. Virus Res 223: 190-196. doi: 10.1016/j.virusres.2016.07.012

Chacón F et al, (2015). The lethality test used for estimating the potency of antivenoms against Bothrops asper snake venom: pathophysiological mechanisms, prophylactic analgesia, and a surrogate in vitro assay. Toxicon 93: 41-50. doi: 10.1016/j.toxicon.2014.11.223

Ma S et al, (2014). Development of a sandwich ELISA for the quantification of enterovirus 71. Cytotechnology 66(3): 413–418. doi: 10.1007%2Fs10616-013-9588-9

Morgeaux S et al, (2013). Validation of a new ELISA method for in vitro potency testing of hepatitis A vaccines. Pharmeur Bio Sci Notes 2013: 64-92. PMID: 24447723

He F et al, (2013). Development of a sensitive and specific epitope-blocking ELISA for universal detection of antibodies to human enterovirus 71 strains. PLoS One  8(1):e55517. doi: 10.1371/journal.pone.0055517

Descamps J et al, (2011). A case study of development, validation, and acceptance of a non-animal method for assessing human vaccine potency. Procedia in Vaccinology  5: 184-191. doi: 10.1016/j.provac.2011.10.018

Poirier B et al, (2010). Would an in vitro ELISA test be a suitable alternative potency method to the in vivo immunogenicity assay commonly used in the context of international Hepatitis A vaccines batch release? Vaccine 28(7): 1796-1802. doi: 10.1016/j.vaccine.2009.12.006

Kumar S et al, (2009). Standardization and validation of Vero cell assay for potency estimation of diphtheria antitoxin serum. Biologicals 37(5): 297-305, doi: 10.1016/j.biologicals.2009.05.002

von Hunolstein C et al, (2008). Evaluation of two serological methods for potency testing of whole cell pertussis vaccines. Pharmeuropa Bio. 2008(1): 7-18. PMID: 19220977

Winsnes R et al, (2006). Collaborative Study for the Validation of Serological Methods for Potency Testing of Diphtheria Toxoid Vaccines - Part 2. Link to article: https://www.edqm.eu/documents/52006/123862/bsp034-p2-dserol.pdf

Shank-Retzlaff M et al, (2005). Correlation between Mouse Potency and In Vitro Relative Potency for Human Papillomavirus Type 16 Virus-Like Particles and Gardasil® Vaccine Samples. Human Vaccines 1(5): 191-197. doi: 10.4161/hv.1.5.2126

Sesardic D et al, (2003). Collaborative Study for the Validation of Serological Methods for Potency Testing of Diphtheria Toxoid Vaccines Extended Study: Correlation of Serology with in vivo Toxin Neutralisation. Link to article: https://www.edqm.eu/documents/52006/123862/bsp034-p1-ext-dserol.pdf

Poirier B et al, (2000). In Vitro Potency Assay for Hepatitis A Vaccines: Development of a Unique Economical Test. Biologicals Volume 28, Issue 4, December 2000, Pages 247-256. 10.1006/biol.2001.0264

Viviani L et al, (2022). Accelerating Global Deletion of the Abnormal Toxicity Test for vaccines and biologicals. Planning common next steps. A workshop Report. Biologicals 78: 17-26. doi: 10.1016/j.biologicals.2022.06.003 

Viviani L et al, (2020). Global harmonization of vaccine testing requirements: Making elimination of the ATT and TABST a concrete global achievement. Biologicals 63: 101-105. doi: 10.1016/j.biologicals.2019.10.007 

Wagner L et al, (2017). Towards replacement of the acellular pertussis vaccine safety test: Comparison of in vitro cytotoxic activity and in vivo activity in mice. Vaccine 35(51): 7160-7165. doi: 10.1016/j.vaccine.2017.10.082

Arciniega J et al, (2016). Alternatives to HIST for acellular pertussis vaccines: progress and challenges in replacement. Pharmeur Bio Sci Notes 2015: 82–96. PMID: 27506225

Dazhi J et al, (2016). Quantitative Detection of Vibrio cholera Toxin by Real-Time and Dynamic Cytotoxicity Monitoring. J Clin Microbiol 51(12) doi: 10.1128/jcm.01959-13

Behrensdorf-Nicol H et al, (2015). "BINACLE" assay for in vitro detection of active tetanus neurotoxin in toxoids. ALTEX 32(2): 137-42. doi: 10.14573/altex.1412181

Xing D et al, (2014). Whole-cell pertussis vaccine potency assays: the Kendrick test and alternative assays. Expert Review of Vaccines 13(10): 1175–1182. doi: 10.1586/14760584.2014.939636

Isbrucker R et al, (2013). Transferability study of CHO cell clustering assays for monitoring of pertussis toxin activity in acellular pertussis vaccines. Pharmeur Bio Sci Notes. 2015: 97-114. PMID: 27506252

Corbel M and Xing D (2004). Toxicity and potency evaluation of pertussis vaccines. Expert Rev Vaccines. 3(1): 89-101. doi: 10.1586/14760584.3.1.89

McKenna A et al, (1995). Attenuated typhoid vaccine Salmonella typhi Ty21a: fingerprinting and quality control. Microbiology 141(8): 1993-2002. doi: 10.1099/13500872-141-8-1993