new software tools beg the question: why dont we monitor
TRANSCRIPT
Protein Metrics SolutionsNew software tools beg the question: Why
don’t we monitor drug product profile directly?
Eric Carlson, President & CEO | Zeta Symposium March 6-8, 2018
[email protected]
BiotechnologyComputing
• Protein analysis should never be limited by instrument software.
• Our software analyzes chromatography and mass spectrometry data in a vendor-neutral manner using best-in-class algorithms.
Zeta Symposium 2018 (March 6-8, 2018) 4SF Chronicle / Brant Ward
Hmm, what’s in there?
Informatics to better design, monitor, and control your process
MANUFACTURING NEWS /JUNE 7, 2016 / GLOBAL PHARMA UPDATE
Zeta Symposium 2018 (March 6-8, 2018) 7
Use data analysis to automatically track and report profile directly…
Deviations
Today’s situation
Timelines are compressing
Start
Launch
Start
Launch
Higher Titers have Increased Challenges in Purification and Production
• Cell lines are being increasingly ‘pushed’ or ‘stressed’
• Purification processes have become longer, more complex, and more expensive
• Product profiles can shift
Source: Bioprocess International, June 2009
Zeta Symposium 2018 (March 6-8, 2018) 10
Genzyme receives PDUFA date from FDA for its Biologics License Application for Lumizyme - 21. January 2010 23:52 http://www.news-medical.net/news/20100121/Genzyme-receives-PDUFA-date-from-FDA-for-its-Biologics-License-Application-for-Lumizyme.aspx
Genzyme makes a very successful enzyme replacement therapy for a rare disease Myosyme
Genzyme built new operation to scale up process from 160L to 2000L
IDENTICAL feed
DIFFERENT
Zeta Symposium 2018 (March 6-8, 2018) 11
Impact of glycosylation
The result Different Critical Quality Attributes (CQAs)! Genzyme had to re-submit as a new product
Regulator Decision:
US: New Clinical Trials EU: Limited Clinical Trials
• ESTIMATED RE-START COST > $40 M USD
• ESTIMATED MARKET SHARE LOSS $200-300M USD
A Real Biotech Story of the need – Genzyme (now Sanofi)
Glycosylation Pattern A Glycosylation Pattern B
A need to understand the design / operationtional space The use of advanced technology (LCMS) would have enabled visibility into operational space deviations
EU: Submission as Biosimilar US: Resubmission for New Product (Pre biosimilar legislation)
Genzyme receives PDUFA date from FDA for its Biologics License Application for Lumizyme - 21. January 2010 23:52 http://www.news-medical.net/news/20100121/Genzyme- receives-PDUFA-date-from-FDA-for-its-Biologics-License- Application-for-Lumizyme.aspx
Zeta Symposium 2018 (March 6-8, 2018) 13
Encouragement of Technology
• ‘Well characterized’ implies detailed data analysis
https://www.landesbioscience.com/journals/mabs/2013MABSED0505.pdf
• Mass spectrometry specifically identified as a tool for characterization… … placing a heavy burden on data analysis
Zeta Symposium 2018 (March 6-8, 2018) 14
Process Considerations - Is This Your Reality?
Offline QA/QC Testing
• Improve speed of transfer
• Better optimize handover from Development to Manufacturing
• Does this warrant change of method?
• Reduced risk & variability
• Waste reduction
Discovery Method
Development Process
Risk Waste
Variability Time
Measure with mass spectrometry
SCIEX has a Strong Track Record with HCP Detection Tracking contaminants and your drug product profile
Examples of monitoring & control points • Indirect attributes
pH Temperature KLa pO2 / pCO2 OD / titer Conductivity
Requiring limited sample prep Intact Mass / Native Mass Spec o Measure drug substance directly o Determine/ provide ratio of major glycoforms o Look for aggregates, changes in clipping, changes in other variants o Near-real time
With some sample prep (enzymatic digest) o Profile HCP, nutrient, and product components to ppm level o Identify contaminants and changes in variants rapidly to low ppm
level
o Product Quality Attributes o Aggregation o Clipped sequence o Glycosylation o Trisulfide bond formation o PTM and sequence variant
Zeta Symposium 2018 (March 6-8, 2018) 16
Limited Sample Prep - Intact Mass & Native Mass Spectrometry
Measurement of the full, un-digested species
Zeta Symposium 2018 (March 6-8, 2018) 17
Opportunity: Downstream Processing for Biopharmaceutical Purity Analysis
0 12 hours
• 40-60 minute sample analysis offline
• Hundreds of liters requires reprocessing
• Initial process recovery yield low
• Final recovery yield after reprocessing around 50-60%
COLLECT WASTEREPROCESS
0 12 hours
Potential now:
reworked
LC-UV-MS System with online software
feedback
Artifact-free deconvolution
Workflow Automation - Intact
Auto-load files Deconvolute &
Workflow Automation - Bispecifics
1 1
Zeta Symposium 2018 (March 6-8, 2018) 23
Use data analysis to automatically track and report profile directly…
Deviations
Some Sample Prep… Peptide-Level Analysis (enzymatic digest and moderate chromatography)
Zeta Symposium 2018 (March 6-8, 2018) 25
Remicade® and RemsimaTM Comparison
N-Glycosylation -Asn300
Remicade 14AO52P1 2017.01
Remicade 14GO43P1 2017.09
Remicade EIM74016P1 2017.08
Remicade EKL97011P1 2017.10
Remsima 12B1C006BA1 2014.12.19
Remsima 12B1C014BA1 2015.05.04
Remsima 12B1C021BA1 2015.12.03
Remsima 12B1C015BA6 2017.05.08
Full profile available….
Change in % a-fucosylation
Infliximab Sample Set 1 Infliximab Sample Set 2
D
FcγR-IIIa Binding is Weaker for Remsima
Run 1 Run 2 Run 3 Run 4 3.2 µM 1.6 µM 0.8µM 0.4 µM
29
R C 1 R C 2 R C 3 R C 4 R S 1 R S 2 R S 3 R S 4
0
K D
Remicade 3.08 ± 0.39 162 ± 18
Remsima 6.15 ± 0.96 351 ± 48
Zeta Symposium 2018 (March 6-8, 2018) 30
Published differences
Host Cell Proteins (HCPs): Unpredictable?
• “The physiochemical properties of the intended recombinant protein … influence the HCPs present at various stages in the bioprocess due to co-purification .... These factors can be quite difficult to predict a priori and are often learned only by testing during process development.”
• .. So it is wise to use a comprehensive approach
Wang et al, Biotechnology and Bioengineering, Vol. 103, No. 3, June 15, 2009
Before there was Mass Spectrometry…
Zeta Symposium 2018 (March 6-8, 2018) 34
Multiple Protein HCP Study (with Bruker)
• Spiked-in 48 proteins at known concentrations
• Ability to quantify down to the ppm level (relative to protein therapeutic)
• Users simply choose the calculation type
• Normalization on a column or row (e.g. within sample, or relative to therapeutic protein)
• Byologic® report for tables and bar charts
• Choice of display formats
Stacked Bar Chart for ‘total’ HCP load
Heatmap table normalized to most abundant protein
within that sample
Automated Calculations in Pivot Table Reports
• Users simply choose the calculation type
• Normalization on a column or row (sample vs protein) can then be used
• Same data can be visualized in multiple ways
(Bruker Maxis)
Compliance
Compliance tools - Usability in routine environments
• LCUV(MS) Peptide mapping: with flags for Out of Tolerance (OOT) • User view for all peaks OOT via a
pre-determined ‘flag’ of peak area
• cGMP: Tools Available • Named user login - implemented
• Automated data storage
Audit Trail
It’s Not Always Forward Planning that Provides Process Improvements
This is the pathway to AbbVie’s front door in late 2016 …
Zeta Symposium 2018 (March 6-8, 2018) 40
It’s Not Always Forward Planning that Provides Process Improvements
And by 2017 things have changed!
Zeta Symposium 2018 (March 6-8, 2018) 41
Testimonials
"With Byologic® I replaced six different analytical LC/MS peptide map methods (six analysts doing six methods) with one MS method. We transferred it to the High Throughput Screening group. They can run that one method faster than it used to take one analyst to run an individual method, and we can generate and ask questions much more quickly.
Using Protein Metrics software, we can also run cell-culture samples directly in days. Prior to implementing their software, it took us six months to send samples to purification and then analyze them."
Matt Traylor, Ph.D., Associate Director of Analytical Development, Shire
"Protein Metrics software is a powerful tool for analyzing glycosylation and other PTMS. Their software greatly alleviates the mass spec analysis bottleneck."
Wilson Phung, Scientific Researcher, Applied Proteomics, Genentech/Roche
“I really like the Protein Metrics software. It helped us to make advances that we may not have been able to with other software. It identified protein impurities that we couldn't make with other software. I was impressed with the auto reporting features and search engine. We realized cost savings and were able to reallocate resources. We can now get deeper into our to data to find what we're looking for. The people are really great to deal with - Eric and the team - fantastic, we get a very good response from them."
Mimi Roy, Ph.D., Director of Analytical Development, BioMarin
Zeta Symposium 2018 (March 6-8, 2018) 42
Thank You
www.proteinmetrics.com [email protected]
Eric Carlson, President & CEO | Zeta Symposium March 6-8, 2018
[email protected]
BiotechnologyComputing
• Protein analysis should never be limited by instrument software.
• Our software analyzes chromatography and mass spectrometry data in a vendor-neutral manner using best-in-class algorithms.
Zeta Symposium 2018 (March 6-8, 2018) 4SF Chronicle / Brant Ward
Hmm, what’s in there?
Informatics to better design, monitor, and control your process
MANUFACTURING NEWS /JUNE 7, 2016 / GLOBAL PHARMA UPDATE
Zeta Symposium 2018 (March 6-8, 2018) 7
Use data analysis to automatically track and report profile directly…
Deviations
Today’s situation
Timelines are compressing
Start
Launch
Start
Launch
Higher Titers have Increased Challenges in Purification and Production
• Cell lines are being increasingly ‘pushed’ or ‘stressed’
• Purification processes have become longer, more complex, and more expensive
• Product profiles can shift
Source: Bioprocess International, June 2009
Zeta Symposium 2018 (March 6-8, 2018) 10
Genzyme receives PDUFA date from FDA for its Biologics License Application for Lumizyme - 21. January 2010 23:52 http://www.news-medical.net/news/20100121/Genzyme-receives-PDUFA-date-from-FDA-for-its-Biologics-License-Application-for-Lumizyme.aspx
Genzyme makes a very successful enzyme replacement therapy for a rare disease Myosyme
Genzyme built new operation to scale up process from 160L to 2000L
IDENTICAL feed
DIFFERENT
Zeta Symposium 2018 (March 6-8, 2018) 11
Impact of glycosylation
The result Different Critical Quality Attributes (CQAs)! Genzyme had to re-submit as a new product
Regulator Decision:
US: New Clinical Trials EU: Limited Clinical Trials
• ESTIMATED RE-START COST > $40 M USD
• ESTIMATED MARKET SHARE LOSS $200-300M USD
A Real Biotech Story of the need – Genzyme (now Sanofi)
Glycosylation Pattern A Glycosylation Pattern B
A need to understand the design / operationtional space The use of advanced technology (LCMS) would have enabled visibility into operational space deviations
EU: Submission as Biosimilar US: Resubmission for New Product (Pre biosimilar legislation)
Genzyme receives PDUFA date from FDA for its Biologics License Application for Lumizyme - 21. January 2010 23:52 http://www.news-medical.net/news/20100121/Genzyme- receives-PDUFA-date-from-FDA-for-its-Biologics-License- Application-for-Lumizyme.aspx
Zeta Symposium 2018 (March 6-8, 2018) 13
Encouragement of Technology
• ‘Well characterized’ implies detailed data analysis
https://www.landesbioscience.com/journals/mabs/2013MABSED0505.pdf
• Mass spectrometry specifically identified as a tool for characterization… … placing a heavy burden on data analysis
Zeta Symposium 2018 (March 6-8, 2018) 14
Process Considerations - Is This Your Reality?
Offline QA/QC Testing
• Improve speed of transfer
• Better optimize handover from Development to Manufacturing
• Does this warrant change of method?
• Reduced risk & variability
• Waste reduction
Discovery Method
Development Process
Risk Waste
Variability Time
Measure with mass spectrometry
SCIEX has a Strong Track Record with HCP Detection Tracking contaminants and your drug product profile
Examples of monitoring & control points • Indirect attributes
pH Temperature KLa pO2 / pCO2 OD / titer Conductivity
Requiring limited sample prep Intact Mass / Native Mass Spec o Measure drug substance directly o Determine/ provide ratio of major glycoforms o Look for aggregates, changes in clipping, changes in other variants o Near-real time
With some sample prep (enzymatic digest) o Profile HCP, nutrient, and product components to ppm level o Identify contaminants and changes in variants rapidly to low ppm
level
o Product Quality Attributes o Aggregation o Clipped sequence o Glycosylation o Trisulfide bond formation o PTM and sequence variant
Zeta Symposium 2018 (March 6-8, 2018) 16
Limited Sample Prep - Intact Mass & Native Mass Spectrometry
Measurement of the full, un-digested species
Zeta Symposium 2018 (March 6-8, 2018) 17
Opportunity: Downstream Processing for Biopharmaceutical Purity Analysis
0 12 hours
• 40-60 minute sample analysis offline
• Hundreds of liters requires reprocessing
• Initial process recovery yield low
• Final recovery yield after reprocessing around 50-60%
COLLECT WASTEREPROCESS
0 12 hours
Potential now:
reworked
LC-UV-MS System with online software
feedback
Artifact-free deconvolution
Workflow Automation - Intact
Auto-load files Deconvolute &
Workflow Automation - Bispecifics
1 1
Zeta Symposium 2018 (March 6-8, 2018) 23
Use data analysis to automatically track and report profile directly…
Deviations
Some Sample Prep… Peptide-Level Analysis (enzymatic digest and moderate chromatography)
Zeta Symposium 2018 (March 6-8, 2018) 25
Remicade® and RemsimaTM Comparison
N-Glycosylation -Asn300
Remicade 14AO52P1 2017.01
Remicade 14GO43P1 2017.09
Remicade EIM74016P1 2017.08
Remicade EKL97011P1 2017.10
Remsima 12B1C006BA1 2014.12.19
Remsima 12B1C014BA1 2015.05.04
Remsima 12B1C021BA1 2015.12.03
Remsima 12B1C015BA6 2017.05.08
Full profile available….
Change in % a-fucosylation
Infliximab Sample Set 1 Infliximab Sample Set 2
D
FcγR-IIIa Binding is Weaker for Remsima
Run 1 Run 2 Run 3 Run 4 3.2 µM 1.6 µM 0.8µM 0.4 µM
29
R C 1 R C 2 R C 3 R C 4 R S 1 R S 2 R S 3 R S 4
0
K D
Remicade 3.08 ± 0.39 162 ± 18
Remsima 6.15 ± 0.96 351 ± 48
Zeta Symposium 2018 (March 6-8, 2018) 30
Published differences
Host Cell Proteins (HCPs): Unpredictable?
• “The physiochemical properties of the intended recombinant protein … influence the HCPs present at various stages in the bioprocess due to co-purification .... These factors can be quite difficult to predict a priori and are often learned only by testing during process development.”
• .. So it is wise to use a comprehensive approach
Wang et al, Biotechnology and Bioengineering, Vol. 103, No. 3, June 15, 2009
Before there was Mass Spectrometry…
Zeta Symposium 2018 (March 6-8, 2018) 34
Multiple Protein HCP Study (with Bruker)
• Spiked-in 48 proteins at known concentrations
• Ability to quantify down to the ppm level (relative to protein therapeutic)
• Users simply choose the calculation type
• Normalization on a column or row (e.g. within sample, or relative to therapeutic protein)
• Byologic® report for tables and bar charts
• Choice of display formats
Stacked Bar Chart for ‘total’ HCP load
Heatmap table normalized to most abundant protein
within that sample
Automated Calculations in Pivot Table Reports
• Users simply choose the calculation type
• Normalization on a column or row (sample vs protein) can then be used
• Same data can be visualized in multiple ways
(Bruker Maxis)
Compliance
Compliance tools - Usability in routine environments
• LCUV(MS) Peptide mapping: with flags for Out of Tolerance (OOT) • User view for all peaks OOT via a
pre-determined ‘flag’ of peak area
• cGMP: Tools Available • Named user login - implemented
• Automated data storage
Audit Trail
It’s Not Always Forward Planning that Provides Process Improvements
This is the pathway to AbbVie’s front door in late 2016 …
Zeta Symposium 2018 (March 6-8, 2018) 40
It’s Not Always Forward Planning that Provides Process Improvements
And by 2017 things have changed!
Zeta Symposium 2018 (March 6-8, 2018) 41
Testimonials
"With Byologic® I replaced six different analytical LC/MS peptide map methods (six analysts doing six methods) with one MS method. We transferred it to the High Throughput Screening group. They can run that one method faster than it used to take one analyst to run an individual method, and we can generate and ask questions much more quickly.
Using Protein Metrics software, we can also run cell-culture samples directly in days. Prior to implementing their software, it took us six months to send samples to purification and then analyze them."
Matt Traylor, Ph.D., Associate Director of Analytical Development, Shire
"Protein Metrics software is a powerful tool for analyzing glycosylation and other PTMS. Their software greatly alleviates the mass spec analysis bottleneck."
Wilson Phung, Scientific Researcher, Applied Proteomics, Genentech/Roche
“I really like the Protein Metrics software. It helped us to make advances that we may not have been able to with other software. It identified protein impurities that we couldn't make with other software. I was impressed with the auto reporting features and search engine. We realized cost savings and were able to reallocate resources. We can now get deeper into our to data to find what we're looking for. The people are really great to deal with - Eric and the team - fantastic, we get a very good response from them."
Mimi Roy, Ph.D., Director of Analytical Development, BioMarin
Zeta Symposium 2018 (March 6-8, 2018) 42
Thank You
www.proteinmetrics.com [email protected]