Airborne Assessment for Aerosols During Manufacturing of Large Molecule Biologics

1. Airborne Assessment for Aerosols During Manufacturing of Large Molecule Biologics AIHCE – Portland, OR Pharma Forum – Wednesday, May 18, 2011 Dave Pearson – Johnson & Johnson PRD

2. Acknowledgements Discussion • Marc Abromovitz, J&J Global Pharma • Brett Chronister and Steve Nowakowski, J&J Malvern EHS • Dave Pearson, J&J Spring House EHS • Kevin Turner, J&J Radnor EHS • Mark Tartaglia, EHS Impact Solutions, LLC

3. J&J Historical Background- IH and Large Molecule Compounds - • Only Qualitative Risk Assessments had been used to evaluate manufacturing processes for large biomolecules • Most large molecule compounds characterized as being low hazard • Not easily absorbed through the skin, digestive system or respiratory tract • Biologics manufacturing processes also characterized as low risk • Primarily wet, closed processes • Consequently, never developed OELs or performed air sampling • More recent decision to apply hazard banding instead of developing OELs for large biomolecules • Interest within EHS to validate these positions

4. How well do large biomolecules (proteins, monoclonal antibodies, etc.) get absorbed? • Not absorbed through the skin • Not likely absorbed through the upper respiratory tract • Due to their large molecular weight, those that reach the deep regions of the lungs have low absorption into the bloodstream: • Insulin (MW = 5,700) approximately 10% absorption • Human Growth Hormone (MW = 30,000) 5% absorption • Typical Biologic (MW > 100,000) 10% absorption conservatively assumed

5. Airborne Exposure to Large Molecule Aerosols vs. Actual Bloodstream Uptake • Inhalable Fraction – The fraction of suspended material in ambient air that actually enters the nose or mouth with the volume of air inhaled (typically up to 100 microns in size). • Respirable Fraction – The fraction of inhalable aerosols that can reach pulmonary region (up to 10 microns in size) • Aerosols greater than 10 microns will often deposit in the nasopharyngeal and tracheobronchial regions. These are trapped in the mucous blanket and removed from the body or swallowed.

6. What are other influences on biological activity for large molecular weight compounds? • Readily break down due to exposure to light, low pH and high temperature • For those compounds that are swallowed, expect compound to be denatured when exposed to acidic environment of the stomach. Therefore, uptake into the bloodstream via the digestive tract is also not expected. • After settling on work surfaces, expect biological activity to be eliminated due to environmental exposure. Therefore, effects of re-suspension in air after drying should be negligible.

7. Purpose of Monitoring • Identify all potential sources of aerosol generation during production of large molecule compounds. • Evaluate methods for potential measurement of employee exposure to large molecule respirable aerosols and compare with lower end of OEL-2 band (20 ug/m3). • Methods considered: • Gravimetric Respirable Aerosol Sampling • Total Protein • Compound-specific IH Method • Real-time Aerosol Monitoring

8. Monitoring Methods Considered -Measuring Total Protein • Advantages • More specific than Total Dust • LLD (100 ng/m3 ) is below the lower end of OEL-2 band • Disadvantages • Does not differentiate between naturally occurring protein • Historically Total Protein IH sampling has not given an accurate picture of true exposure

9. Monitoring Methods Considered -Specific IH Method • Advantages • Most accurate way to determine airborne employee exposure levels • Compound-specific • Disadvantages • Compound-specific, therefore need new method for each molecule • Delayed time to receive results

10. Monitoring Methods Considered -Real-Time Aerosol Monitoring • Advantages • Can provide accurate real time data on respirable aerosols • Can be used at multiple sites for multiple products • Can immediately identify activities and locations of leakage/release • Disadvantages • Not validated for real-time measurement of inhalable aerosols >10 microns • Does not differentiate aerosol that is not part of the process. Need to assume that the detected aerosol is 100% product. • Not a personal sample

11. Monitoring Strategy For Real-Time Aerosol Monitoring • Real-time monitoring conducted at two (2) of the J&J Biologics production facilities. • Malvern – GMP, commercial product facility • Spring House – Pilot scale, clinical product facility • Area readings taken at workstation operator breathing zones and at the point source for tasks that have the highest potential for generating aerosols. • Work area background samples taken during non-processing periods for comparison purposes.

12. Real Time Aerosol Monitor • DustTrak™ II Model 8530 Aerosol Monitor (TSI, Inc.) • Desktop and Handheld models • Battery operated • Light scattering LASER photometer • Simultaneous measurement of mass and size fraction • Data logger Desktop Handheld

13. Potential Aerosolization Points in Manufacturing • Releases during equipment purging or in-process sampling • Leaks at connections, hoses, tri-clamps, etc. • Final Fill Process (product is dispensed into bulk containers) • Equipment Disassembly

14. Typical Monoclonal Antibody Bulk Process s s Recovery 2L - 20L Clarification AffinityChromatography Harvest (ATF) Cell Bank Pre-Culture Bioreactor = Sampling Location s Media Prep Buffer Prep s s s Final Filtration Cation ExchangeChromatography Anion ExchangeChromatography Virus Filtration Step Bulk Product VirusInactivation

15. Examples of processes with potential for aerosolization

16. Air Sampling Results – GMP Operations Data analysis performed using IHDataAnalyst, V1.05, Exposure Assessment Solutions, Inc.

17. Air Sampling Results – nonGMP Operations Data analysis performed using IHDataAnalyst, V1.05, Exposure Assessment Solutions, Inc.

18. Conclusions Discussion • 95% UCL for both Respirable and Total aerosol fraction sampling results were one-half or less lower end of OEL-2 band [20 ug/m3] without adjusting for % Active Ingredient content. • No operations were identified that require additional engineering controls or use of respirators per J&J criteria. • Respirable aerosol sampling results were similar to Background results (substantial overlap in 95% CL band). • Respirable aerosol sampling results for GMP operations were similar to non-GMP operations (non-GMP 95% CL band contained within GMP 95% CL band).

19. Conclusions (cont.) Discussion • DustTrak useful as survey instrument to provide real-time concentrations for Respirable fraction during large biomolecule processing. • Handheld unit may provide more flexibility for field work. • Clean room environment aided in ability to use non-selective instrument to measure target aerosols.

20. Next Steps Discussion • Complete additional comparative statistical analysis of data • Respirable data sets vs. Background sets • GMP data set vs. non-GMP set • Collect additional data at PA facilities • Collect data at other J&J Biologics facilities • St. Louis • Leiden (The Netherlands) • Cork (Ireland)

21. ? QUESTIONS ?

22. Respirator Decision Analysis (Respirable GMP - No LOD) DATA

23. Respirator Decision Analysis (Respirable GMP - LOD) DATA

24. References Discussion • References: • Model 8530/8531/8532,DUSTTRAK™ II Aerosol Monitor Operation and Service Manual; TSI Incorporated, Revision E, March 2010 • “Counting and particle transmission efficiency of the aerodynamic particle sizer”; J. Volckens, T.M. Peters / Aerosol Science 36 (2005) 1400–1408

25. Discussion Discussion • DustTrak II Instrument • Calibration • Factory calibration maintained • Factory calibration factor used • Specific aerosol calibration factor not determinable due to aqueous nature • Instrument design not include accelerating nozzle that has been reported to underestimate liquid aerosols in other instruments [J. Volckens, T.M. Peters / Aerosol Science 36 (2005) 1400–1408] • Maintenance performed per Operation Manual. • Desktop model difficult to handle for field work due to size/weight. • Product Formulations • Range of typical Active Ingredient % concentration is up to 5% • Results were not adjusted to account for AI concentration.

26. Summary of Benchmarking • Data collected early 2010 • Limited number of pharmaceutical organizations surveyed • Two (2) companies have developed compound-specific IH methods with contract IH lab (compounds were Carcinogenic or Cytotoxic). Both companies sampled stages in the process that they believed presented the greatest risk of employee exposure. Both companies reported results in low ng/m3 or ND. • Three (3) companies currently relying on qualitative risk assessments. They believe qualitative risk assessments are adequate due to the nature of the compounds and the processes.

27. Conclusions (Alternate) Discussion • 95% UCL for both Respirable and Total aerosol fraction sampling results were one-half or less lower end of OEL-2 band [20 ug/m3] without adjusting for % Active Ingredient content. • No operations were identified that require engineering controls or use of respirators per J&J criteria <INSERT CRITERIA> . • Comparative aerosol sampling results between Respirable fraction and Background results were <INSERT CONCLUSION> . • Comparative aerosol sampling results between GMP and non-GMP operations were <INSERT CONCLUSION> .