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
Acknowledgements
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
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, no longer develop 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
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
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.
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.
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
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
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
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
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.
Real Time Aerosol Monitor
DustTrak™ II Model 8530
Aerosol Monitor (TSI, Inc.)
Desktop
Handheld
Desktop and Handheld models
Battery operated
Light scattering LASER
photometer
Simultaneous measurement of
mass and size fraction
Data logger
Potential Aerosolization Points
in Manufacturing
Releases during equipment purging or inprocess sampling
Leaks at connections, hoses, tri-clamps, etc.
Final Fill Process (product is dispensed into bulk
containers)
Equipment Disassembly
Typical Monoclonal Antibody Bulk Process
s
s
Recovery
2L - 20L
Cell Bank
Pre-Culture
Bioreactor
Harvest
(ATF)
Clarification
Affinity
Chromatography
s = Sampling Location
Media Prep
Buffer Prep
s
Bulk
Product
Final
Filtration
s
s
Cation
Anion
Virus
Virus
Exchange
Filtration Step Exchange
Inactivation
Chromatography Chromatography
Examples of processes with
potential for aerosolization
Air Sampling Results – GMP Operations
GMP Respirable
GMP Total
GMP Background
ORDER STATISTICS
ORDER STATISTICS
ORDER STATISTICS
n=
17
n=
17
n=
10
min =
1.0 ug/m3
min =
1.0 ug/m3
min =
1.0 ug/m3
max =
10 ug/m3
max =
10 ug/m3
max =
5.0 ug/m3
median =
1.0 ug/m3
median =
1.0 ug/m3
median =
1.0 ug/m3
DESCRIPTIVE STATISTICS
DESCRIPTIVE STATISTICS
DESCRIPTIVE STATISTICS
mean =
2.3 ug/m3
mean =
1.8 ug/m3
mean =
1.4 ug/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
X0.95 =
5.7
95%LCL =
3.9
COMPLIANCE STATISTICS
95%UCL
= 11
X0.95 =
4.0
95%LCL =
2.8
COMPLIANCE STATISTICS
95%UCL
=6.9
Data analysis performed using IHDataAnalyst, V1.05, Exposure Assessment Solutions, Inc.
X0.95 =
2.7
95%LCL = 95%UCL =
2.0
5.2
Air Sampling Results – nonGMP Operations
Non-GMP Total
Non-GMP Respirable
Non-GMP Background
ORDER STATISTICS
ORDER STATISTICS
ORDER STATISTICS
n=
n=
n=
7
7
2
min =
5.0 ug/m3
min =
4.0 ug/m3
min =
3.0 ug/m3
max =
6.0 ug/m3
max =
5.0 ug/m3
max =
9.0 ug/m3
median =
5.0 ug/m3
median =
4.0 ug/m3
median =
6.0 ug/m3
DESCRIPTIVE STATISTICS
DESCRIPTIVE STATISTICS
DESCRIPTIVE STATISTICS
mean =
5.4 ug/m3
mean =
4.3 ug/m3
mean =
6.0 ug/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
X0.95 =
6.3
95%LCL = 95%UCL =
5.9
7.5
COMPLIANCE STATISTICS
X0.95 =
5.1
95%LCL =
4.7
COMPLIANCE STATISTICS
95%UCL
= 6.2
Data analysis performed using IHDataAnalyst, V1.05, Exposure Assessment Solutions, Inc.
X0.95 =
19
95%LCL = 95%UCL =
75 3.76E0006
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).
Conclusions
Discussion (cont.)
 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 nonselective instrument to measure target aerosols.
Discussion
Next Steps
 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
 Leiden (The Netherlands)
 Cork (Ireland)
? QUESTIONS ?