Animal use in biologics development, production, and testing - PowerPoint PPT Presentation

Animal use in biologics development, production, and testing Implementing Nonanimal Approaches to Human and Veterinary Vaccine Testing: Achieving Scientific and Regulatory Success for Rabies and Beyond October 10, 2018 Jeffrey Brown JeffreyB@PISCLtd.org.uk

PETA International Science Consortium 2

PETA International Science Consortium See full list of publications at PISCLtd.org.uk/SciPubs 3

Outline 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 4

2011 rabies vaccine workshop 5

2011 rabies vaccine workshop 6

2011 rabies vaccine workshop: NIH test problems 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 7 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 8 Bruckner 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 9 Bruckner 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.

2011 rabies vaccine workshop: Progress 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 challenge o 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.) 10 Stokes 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.

2011 rabies vaccine workshop: Progress 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 challenge o 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.) 11 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.

Problems: counting animals 12

Problems: counting animals 13 Laboratory Animal Welfare Act of 1966, P.L. 89-544

Problems: Not just the NIH test 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 14

Problems: Animal-derived antibodies 15 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.

Problems: Animal-derived antibodies 16 Groff K, Brown J, Clippinger AJ. Modern affinity reagents: recombinant antibodies and aptamers. Biotechnol Adv . 2015;33(8):1787-1798.

Problems: Animal-derived antibodies 17 Image: PETA US

Progress to come: recombinant antibodies o Phage display 18

Progress to come: recombinant antibodies 19 Bradbury A, Plückthun A. Reproducibility: Standardize antibodies used in research. Nature . 2015;518(7537):27-29.

Progress to come: recombinant antibodies 20 Bradbury A, Plückthun A. Reproducibility: Standardize antibodies used in research. Nature . 2015;518(7537):27-29.

Progress to come: recombinant antibodies 21

Progress to come: recombinant antibodies 22

Animal use: antibody sources 23 Images: USDA Inspection Reports

Animal use: antibody sources 24 Images: USDA Inspection Reports

Animal use: antibody sources 25 Images: USDA Inspection Reports

Animal use: antibody sources 26 Images: USDA Inspection Reports

Animal use: antibody sources 27 Images: USDA Inspection Reports

Animal use: antibody sources 28 Images: PETA India

Animal use: antibody sources 29 Images: PETA India

Animal use: antibody sources 30 Images: PETA India

Progress to come: replacing animal antibodies 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 31

Progress to come: pyrogen testing 32 Images: Cruelty Free International; Timothy Fadek / Corbis / Getty; The Atlantic

Progress to come: pyrogen testing 33 Zhang S. The Last Days of the Blue-Blood Harvest. The Atlantic . May 9, 2018.

Progress to come: pyrogen testing 34 Image: Merck

Summary o Success is difficult to quantify o How to measure impact of animal replacement, reduction, refinement policies? o NIH test replacement is within reach o Improved antibody sources should be prioritised o Antibody production → recombinant methods Contact: o Research, diagnostic, and therapeutic relevance Jeffrey Brown JeffreyB@PISCLtd.org.uk o Ensuring use of other available replacements www.PISCLtd.org.uk o RPT/BET → MAT @PISCLtd 35