Animal use in biologics development, production, and testingnonanimal-approaches-to-vaccine-testing.iabs.org/... · • Pyrogen testing 4 Outline. 2011 rabies vaccine workshop 5.

Animal use in biologics development, production, and testingImplementing Nonanimal Approaches to Human and Veterinary Vaccine Testing: Achieving Scientific and Regulatory Success for Rabies and Beyond

October 10, 2018

Jeffrey BrownJeffreyB@PISCLtd.org.uk

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PETA International Science Consortium

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PETA International Science Consortium

See full list of publications at PISCLtd.org.uk/SciPubs

o Progress and problems• 3Rs policies for rabies vaccines (etc.)• Tracking animal use and 3Rs policy implementation• “Peripheral” animal use

o Progress to come• Antibody production• Pyrogen testing

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Outline

2011 rabies vaccine workshop

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2011 rabies vaccine workshop

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2011 rabies vaccine workshop: NIH test problems

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o ~200 mice per test

o ~50% experience significant unrelieved pain prior to euthanasia

o Up to 42% of tests invalid, must be repeated

o Up to 400% differences in estimated potency

Stokes W, McFarland R, Kulpa-Eddy J, Gatewood D, Levis R, Halder M, Pulle G, Kojima H, Casey W, Gaydamka A, Miller T, Brown K, Lewis C, Capsal JM, Bruckner L, Gairola S, Kamphuis E, Rupprecht CE, Wunderli P, McElhinney L, De Mattia F, Gamoh K, Hill R, Reed D, Doelling V, Johnson N, Allen D, Rinkel L, Jones B. Report on the international workshop on alternative methods for human and veterinary rabies vaccine testing: state of the science and planning the way forward. Biologicals. 2012;40(5):369-381.

NIH test

8Bruckner L, Cussler K, Halder M, Barrat J, Castle P, Duchow K, Gatewood DM, Gibert R, Groen J, Knapp B, Levis R, Milne C, Parker S, Stünkel K, Visser N, Volkers P; ECVAM. Three Rs approaches in the quality control of inactivated rabies vaccines. The report and recommendations of ECVAM workshop 48. Altern Lab Anim. 2003; 31(4):429-454.

NIH test

9Bruckner L, Cussler K, Halder M, Barrat J, Castle P, Duchow K, Gatewood DM, Gibert R, Groen J, Knapp B, Levis R, Milne C, Parker S, Stünkel K, Visser N, Volkers P; ECVAM. Three Rs approaches in the quality control of inactivated rabies vaccines. The report and recommendations of ECVAM workshop 48. Altern Lab Anim. 2003; 31(4):429-454.

o Implementation strategy to:o Replace NIH test with ELISA and/or SNT

• EPAA pre-validation study and EDQM international collaborative study

o Reduce numbers of mice used to meet NIH test requirements• USDA Center for Veterinary Biologics (CVB) Notice 13-10,

eliminates upper limit LD50 for a valid challengeo Reduce pain for mice used in NIH test

• USDA SAM 308 revised, allows anesthesia for mice prior to virus injection

• USDA CVB Notice 12-12, firms encouraged to use anesthesia prior to virus injection (etc.)

2011 rabies vaccine workshop: Progress

10Stokes W, McFarlandR, Kulpa-Eddy J, Gatewood D, Levis R, Halder M, Pulle G, Kojima H, Casey W, Gaydamka A, Miller T, Brown K, Lewis C, Capsal JM, Bruckner L, Gairola S, Kamphuis E, Rupprecht CE, Wunderli P, McElhinney L, De Mattia F, Gamoh K, Hill R, Reed D, Doelling V, Johnson N, Allen D, Rinkel L, Jones B. Report on the international workshop on alternative methods for human and veterinary rabies vaccine testing: state of the science and planning the way forward. Biologicals. 2012;40(5):369-381.

o Implementation strategy to:o Replace NIH test with ELISA and/or SNT

• EPAA pre-validation study and EDQM international collaborative study

o Reduce numbers of mice used to meet NIH test requirements• USDA Center for Veterinary Biologics (CVB) Notice 13-10,

eliminates upper limit LD50 for a valid challengeo Reduce pain for mice used in NIH test

• USDA SAM 308 revised, allows anesthesia for mice prior to virus injection

• USDA CVB Notice 12-12, firms encouraged to use anesthesia prior to virus injection (etc.)

2011 rabies vaccine workshop: Progress

11Stokes W, McFarland R, Kulpa-Eddy J, Gatewood D, Levis R, Halder M, Pulle G, Kojima H, Casey W, Gaydamka A, Miller T, Brown K, Lewis C, Capsal JM, Bruckner L, Gairola S, Kamphuis E, Rupprecht CE, Wunderli P, McElhinney L, De Mattia F, Gamoh K, Hill R, Reed D, Doelling V, Johnson N, Allen D, Rinkel L, Jones B. Report on the international workshop on alternative methods for human and veterinary rabies vaccine testing: state of the science and planning the way forward. Biologicals. 2012;40(5):369-381.

Problems: counting animals

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Problems: counting animals

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Laboratory Animal Welfare Act of 1966, P.L. 89-544

o Required to carry out NIH test• Virus propagation: birds, mice, hamsters

o Required to meet product standard requirements• Immunogenicity: target species• Purity: mice• Safety: mice, rabbits, other species• Duration of immunity: target species

o “Peripheral” animals used in experiments• Antibody sources• Serum sources• Pyrogen testing

Problems: Not just the NIH test

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Problems: Animal-derived antibodies

15Stokes W, McFarland R, Kulpa-Eddy J, Gatewood D, Levis R, Halder M, Pulle G, Kojima H, Casey W, Gaydamka A, Miller T, Brown K, Lewis C, Capsal JM, Bruckner L, Gairola S, Kamphuis E, Rupprecht CE, Wunderli P, McElhinney L, De Mattia F, Gamoh K, Hill R, Reed D, Doelling V, Johnson N, Allen D, Rinkel L, Jones B. Report on the international workshop on alternative methods for human and veterinary rabies vaccine testing: state of the science and planning the way forward. Biologicals. 2012;40(5):369-381.

Problems: Animal-derived antibodies

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Groff K, Brown J, Clippinger AJ. Modern affinity reagents: recombinant antibodies and aptamers. Biotechnol Adv. 2015;33(8):1787-1798.

Problems: Animal-derived antibodies

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Image: PETA US

Progress to come: recombinant antibodies

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o Phage display

Progress to come: recombinant antibodies

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Bradbury A, Plückthun A. Reproducibility: Standardize antibodies used in research. Nature. 2015;518(7537):27-29.

Progress to come: recombinant antibodies

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Bradbury A, Plückthun A. Reproducibility: Standardize antibodies used in research. Nature. 2015;518(7537):27-29.

Progress to come: recombinant antibodies

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Progress to come: recombinant antibodies

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Animal use: antibody sources

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Images: USDA Inspection Reports

Animal use: antibody sources

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Images: USDA Inspection Reports

Animal use: antibody sources

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Images: USDA Inspection Reports

Animal use: antibody sources

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Images: USDA Inspection Reports

Animal use: antibody sources

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Images: USDA Inspection Reports

Animal use: antibody sources

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Images: PETA India

Animal use: antibody sources

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Animal use: antibody sources

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Images: PETA India

Progress to come: replacing animal antibodies

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o Datmab project• Phage display, select recombinant human antibodies against

diphtheria toxin• Assess toxin neutralization of toxin binding antibodies• Convert neutralizing antibodies with best binding affinity to IgG • Freeze-dry formulation and stability testing • Public health stakeholders, further development

Progress to come: pyrogen testing

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Images: Cruelty Free International; Timothy Fadek / Corbis / Getty; The Atlantic

Progress to come: pyrogen testing

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Zhang S. The Last Days of the Blue-Blood Harvest. The Atlantic. May 9, 2018.

Progress to come: pyrogen testing

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Image: Merck

Summary

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o Success is difficult to quantifyo How to measure impact of animal replacement,

reduction, refinement policies?

o NIH test replacement is within reacho Improved antibody sources should be prioritised

o Antibody production → recombinant methodso Research, diagnostic, and therapeutic relevance

o Ensuring use of other available replacementso RPT/BET → MAT

Contact: Jeffrey Brown

JeffreyB@PISCLtd.org.ukwww.PISCLtd.org.uk

@PISCLtd