Airborne Assessment for Aerosols During Manufacturing

of Large Molecule Biologics

AIHCE – Portland, OR

Pharma Forum – Wednesday, May 18, 2011

Dave Pearson – Johnson & Johnson PRD

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

Acknowledgements

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

J&J Historical Background- IH and Large Molecule Compounds -

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

How well do large biomolecules (proteins, monoclonal antibodies, etc.) get absorbed?

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.

Airborne Exposure to Large Molecule Aerosols vs. Actual Bloodstream Uptake

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.

What are other influences on biological activity for large molecular weight compounds?

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

Purpose of Monitoring

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

Monitoring Methods Considered -Measuring Total Protein

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

Monitoring Methods Considered -Specific IH Method

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

Monitoring Methods Considered -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.

Monitoring Strategy For Real-Time Aerosol Monitoring

DustTrak™ II Model 8530 Aerosol Monitor (TSI, Inc.)Desktop and Handheld modelsBattery operatedLight scattering LASER photometerSimultaneous measurement of mass and size fractionData logger

Real Time Aerosol Monitor

Desktop Handheld

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

Potential Aerosolization Points in Manufacturing

Typical Monoclonal Antibody Bulk Process

AffinityChromatography

2L - 20L

BioreactorPre-CultureCell BankClarificationHarvest

(ATF)

Recovery

VirusInactivation

VirusFiltration Step

Anion Exchange

Chromatography

Cation Exchange

Chromatography

BulkProduct

FinalFiltration

Media Prep

Buffer Prep

s s

ss

s

s = Sampling Location

Examples of processes with potential for aerosolization

Air Sampling Results – GMP Operations

GMP Total GMP Respirable GMP Background

ORDER STATISTICS ORDER STATISTICS ORDER STATISTICS  

n = 17 n = 17 n = 10  

min = 1.0ug/m3 min = 1.0ug/m3 min = 1.0ug/m3  

max = 10ug/m3 max = 10ug/m3 max = 5.0ug/m3  

median = 1.0ug/m3 median = 1.0ug/m3 median = 1.0ug/m3  

DESCRIPTIVE STATISTICS DESCRIPTIVE STATISTICS DESCRIPTIVE STATISTICS  

mean = 2.3ug/m3 mean = 1.8ug/m3 mean = 1.4ug/m3  

sd = 2.4 sd = 2.3 sd = 1.3  

gm = 1.6 gm = 1.3 gm = 1.2  

gsd = 2.14 gsd = 1.95 gsd = 1.66  

COMPLIANCE STATISTICS COMPLIANCE STATISTICS COMPLIANCE STATISTICS  

X0.95 = 5.795%LCL =

3.995%UCL =

11 X0.95 = 4.095%LCL =

2.895%UCL

=6.9 X0.95 = 2.795%LCL =

2.095%UCL =

5.2

Data analysis performed using IHDataAnalyst, V1.05, Exposure Assessment Solutions, Inc.

Air Sampling Results – nonGMP Operations

Non-GMP Total Non-GMP Respirable Non-GMP Background

ORDER STATISTICS ORDER STATISTICS ORDER STATISTICS  

n = 7 n = 7 n = 2  

min = 5.0ug/m3 min = 4.0ug/m3 min = 3.0ug/m3  

max = 6.0ug/m3 max = 5.0ug/m3 max = 9.0ug/m3  

median = 5.0ug/m3 median = 4.0ug/m3 median = 6.0ug/m3  

DESCRIPTIVE STATISTICS DESCRIPTIVE STATISTICS DESCRIPTIVE STATISTICS  

mean = 5.4ug/m3 mean = 4.3ug/m3 mean = 6.0ug/m3  

sd = 5.0 sd = 5.0 sd = 4.2  

gm = 5.4 gm = 4.3 gm = 5.2  

gsd = 1.1 gsd = 1.12 gsd = 2.17  

COMPLIANCE STATISTICS COMPLIANCE STATISTICS COMPLIANCE STATISTICS  

X0.95 = 6.395%LCL =

5.995%UCL =

7.5 X0.95 = 5.195%LCL =

4.795%UCL =

6.2 X0.95 = 1995%LCL =

7595%UCL = 3.76E0006

Data analysis performed using IHDataAnalyst, V1.05, Exposure Assessment Solutions, Inc.

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).

Conclusions

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.

Conclusions (cont.)

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)

Next Steps

? QUESTIONS ?

07.14.2010 22

Sample # Conc LOD

16 0.001  

19 0.004  

21 0.001  

25 0.003  

26 0.001  

27 0.001  

28 0.001  

29 0.001  

30 0.001  

35 0.001  

37 0.001  

39 0.001  

41 0.001  

42 0.01  

43 0.001  

45 0.001  

47 0.001  

DATA

Respirator Decision Analysis (Respirable GMP - No LOD)

07.14.2010 23

Sample # Conc LOD

16 0.001  

19 0.004  

21 0.001  Y

25 0.003  

26 0.001  Y

27 0.001  Y

28 0.001  Y

29 0.001  Y

30 0.001  Y

35 0.001  Y

37 0.001  

39 0.001  Y

41 0.001  Y

42 0.01  

43 0.001  Y

45 0.001  Y

47 0.001  Y

DATA

Respirator Decision Analysis (Respirable GMP - LOD)

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

References

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.

Discussion

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.

Summary of Benchmarking

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> .

Conclusions (Alternate)