Target 3-5M doses
Presentation Outline Bringing a vaccine from the 40’s into modern biotech Protein Sciences BEVS technology and advantages
Influenza vaccine challenges and our strategy to overcome
Some background on influenza and influenza vaccines How we address influenza “constant change” through the product life cycle Development, Commercial production and Production expansion
Final thoughts and acknowledgement 2
Meriden, CT FDA licensed, multiproduct facilities 50L & 600L up to 500K doses
Pearl River, NY (formerly Pfizer) FDA licensed Flublok & Panblok 2,000L ~ 5M doses Gifu, Japan (UNIGEN) cGMP 2x21,000L >30M doses Expanding to 6x21,000L 3
Sf9 cells
SF+ cells
Sf9 cells cloned in 1983 from IPLB-Sf-21AE cells (isolated in 1970) Grown in fetal bovine serum Deposited at ATCC in 1985 by Texas A&M University (Cherry/Smith)
At PSC
Selective pressure in serum-free media in 1993 Unique phenotypic and genotypic properties Produce high titer baculovirus Patented Licensed for used in commercial production of USDA licensed veterinary vaccine and human products (US/Europe)
Autographa californica (Alfalfa Looper) Virus AcMNPV, E2 isolate AcMNPV polyhedra were isolated/cloned from a single-fieldcollected alfalfa looper larva Introduced restriction sites to facilitate generation and cloning of recombinant viruses
At PSC Infected our expresSF+ with parental, modified AcMNPV Harvest the virus rich supernatant, frozen, qualified, and designated Master BaculoVirus Bank (MVB)
DNA is isolated from MVB, linearized and recombined with transfer plasmid to form Working Virus Banks
Versatility
Multiproduct facility design
Speed
Single robust cell line for all products
Maximize capacity utilization Cloning in 21 days
Reliable scale-up
Current scale 2,000L, partners scale up to 21,000L
Low cost
High yields, low-cost proprietary media
Safety
BV highly specific to insect cells
GMP Production
Process optimization for new antigens High-density fermentation
No handling of dangerous virus
Concentrated DS
No containment required 6
Influenza vaccine challenges and our strategy to overcome Addressing “constant change” through the Product life cycle
7
Major surface glycoprotein, coat of the influenza virus, Antibodies against HA protect against influenza – the primary active ingredient in licensed vaccine
Changes in HA lead to annual epidemics requiring annual update of vaccine WHO /VRBPAC select the strains annually for the vaccine manufacturing 8
Typically we can expect at least one vaccine strain to change with each season FDA Selected Vaccine Strains by Season Season
H1N1 strain
H3N2 strain
B strain
2015-16
A/California/07
A/Switzerland
B/Phuket
2014-15
A/California/07
A/Texas
B/Massachusetts/2
2013-14
A/California/07
A/Texas
B/Massachusetts/2
2012-13
A/California/07
A/Victoria/361
B/Wisconsin/1
2011-12
A/California/07
A/Perth/16
B/Brisbane/60
2010-11
A/California/07
A/Perth/16
B/Brisbane/60
2009-10
A/Brisbane/59
A/Brisbane/10
B/Brisbane/60
2008-09
A/Brisbane/59
A/Brisbane/10
B/Florida/04
2007-08
A/Solomon Islands/03
A/Wisconsin/67
B/Malaysia/2506
2006-07
A/New Caledonia/20
A/Wisconsin/67
B/Malaysia/2506
Traditional Production • • •
Grow pathogen (in eggs) Inactivate pathogen Purify antigen (i.e., the active ingredient)
Challenges • Long production cycle • Affected by avian influenza outbreaks • Egg adaptation of virus • Egg allergies
Cell Culture Vaccines • Cell growth • MDCK, Vero, PER.C6 • Infection, virus generation • Inactivation / Splitting • Formulate vaccine
Challenges • High level of containment • Adaptation of virus to new substrate
Challenges overcome by recombinant HA vaccine manufacture
Engineer BV with gene of interest Hemagglutinin
BV highly specific to insect cells Powerful promoter generates high yields
Insect cells are cultured in fermentors
Infect cells with engineered virus
Incubate infection for ~48 - 72 hours
Purify protein to >90% Protein forms rosettes Formulate with PBS into vaccine
Flublok® 2013 Approval - Validation of the Platform 11
Trivalent Recombinant HA (rHA) Antigen Vaccine Produced in vitro via insect cell culture/baculovirus technology platform HA matches wild type human virus, no egg-drift 3X active ingredient compared to regular egg vaccine
Flublok rHA Antigens Highly purified, typical purity >90% Correct 3-D structure, biologically active Hemagglutination activity Induce protective immune responses HA antibodies/surrogate marker of efficacy Neutralizing antibodies
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 600L
• Universal Process • Ready to optimize for strain changes
• Yield Increase 10X • Cadence 1.75 batch/week • Formulation, delivery
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand 13
• Four (4) weeks to GMP production • Universal Process • Ready to optimize for strain changes
Challenges for an Influenza Vaccine
• Target 30M doses
Platform Technology
Next Generation Process
Seasonal Strains – Trivalent and quadrivalent formulation Drug Substance Changes - rHA protein changes constantly and the process must accommodate these changes Potency assay reagents - provided by CBER for the season, time critical Annual License Approval – Supplemental BLA submitted annually in time to release, pack and distribute product by early 3rd Q
• Yield Increase 10X
• Increased cadence
From a Process Development Perspective Robust, reproducible process – Universal process, flexible and adaptable Well understood and controlled – Deliberately designed operational spaces to allow optimization for antigen changes Routine, proceduralized approach - Complete optimization, if required in an extremely short time window
• Four (4) weeks to GMP production • Universal Process • Ready to optimize
2. infect
Disc Stack Centrifuge
Cell Pellet
for strain changes
Platform Technology
Depth • Target 30M doses Filtration
Next Generation Process
• Yield Increase 10X • Increased cadence
1. seed
Protein Production
Harvest
Extraction
Clarification
Purification Capture
Purify
DNA removal
Ultrafiltration/Formulation
• Four (4) weeks to GMP production • Universal Process • Ready to optimize for strain changes
Surveillance and HA Sequence Analysis
Cloning of Influenza HA Genes into Transfer Vector
• Data analysis : SH change, CDC, WHO, IFPMA, etc. • Year around HA sequences analysis • 1st option – Cloning from influenza virus • 2nd option – Site Directed Mutagenesis of previously generated HA genes • 3d option – Gene synthesis
Strain Selection
• Target 30M doses
Platform Technology
Next Generation Process
• Yield Increase 10X
• Increased cadence
3 Days
Transfer Vector Insect Cell Transfection and Plaque Purification
Baculovirus Bank Generation
• Co-transfection transfer vector with MVS baculovirus linear DNA • Plaque purification and screening
• Minimum passages during virus scale-up • Stable in liquid N2 • Quality tests ; PCR, Western, SDS-PAGE, etc.
21 Days
Working Virus Bank
• Four (4) weeks to GMP production • Universal Process • Ready to optimize for strain changes
Step
Isotype variability (H1, H3, B)
Cell Culture
upstream
Protein Production
Next Generation Process
Variable • Yield Increase 10X parameters • Target 30M doses
• Increased cadence
universal yes
Harvest
downstream
Seasonal strain variability
Platform Technology
Infrequent
Cell viability at harvest
no
Extraction
yes
Infrequent
pH; [detergent]
Clarification
yes
no
DF Filter type
IEX
yes
Sometimes
pH; [salt]; column type; [detergent]
HIC
yes
Sometimes
pH; [salt]; [detergent]
DNA Removal
Universal
Ultrafiltration Formulation
Universal
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Efficient but
Platform Technology
Framed by the design space for each unit operation • Target 30M doses
• Yield Increase 10X
Next Generation Process
Product Launch
limited supply
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years
• Fully contained within the ranges of the validated process • Focus on updates to specific set-points, key requirements Meet established process performance Maintain quality attributes and stability profile Optimize yield • Driven by known changes in the rHA protein and process knowledge • Sometimes, no changes are required specially for rHA of same lineage • Increased cadence
licensed process
• Capacity to expand
Proceduralized approach – SOP driven work • Rapid assessment with a qualified scale-down model or at production scale • Clearly defined acceptance criteria determines need for optimization • Changes are implementation following Quality System and do no require regulatory approval • Developed in collaboration with the FDA
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Efficient but
Platform Technology
Product Launch
limited supply
Large 2013 - FDA approves Flublok for the prevention of influenza Scale Manuf in adults 18 - 49 years old • Target 30M doses
Next Generation Process
• Yield Increase 10X
• Increased cadence
.
• Target 3-5M doses
• Fast, 1-2 years licensed process
“The Evolution, and Revolution, of Flu Vaccines”
• Capacity to expand
http://www.fda.gov/ForConsumers/ConsumerUpdates/ucm336267.htm
First recombinant influenza vaccine The pandemic solution Only pandemic vaccine that can be quickly manufactured and/or transferred and manufactured in other countries
2014 - Flublok approved for adults 18 and older 2015 - Large Scale Manufacturing approved for production of Flublok and Panblok 19
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Target 30M doses • Yield Increase 10X • Increased cadence
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
20
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Yield Increase 10X • Cadence 1.75 batch/week • Formulation, delivery
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand 21
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
CMO - Expedient, costly, restricted
Greenfield Facility: Perfect Fit, high cost, long time
Adapt an Existing Facility: Challenging, but affordable & flexible •
Relatively minor expenses Ready for capacity expansion, Attractive lease and government support
Moved-in Dec. 3rd 2012; Approved May 2015! 22
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Process Design
Readiness
~ 1 month BARDA Proposal
Preparation for Lease agreement
Match or “Fit” the Process to facility & equipment
Scope, quote and ready for equipment instrument purchase
Define high level design As-Is, Adapt, New Gap Assessment Characterization vs. Design
Risk Analysis Gap Closure Program
Focus: efficiency, cost and short lead times Raw materials & components plan media, resins, etc.
Start-Up & Confirmation 100 days! 1st batch March 26 2013 Match process parameters, yield, purity Optimize & complete documentation
Move to qualification & preparation for PV 23
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Upstream Process & Equipment Bacterial fermentation train retrofitted for cell culture Install oxygen service, new impellers, spargers, and DO control system Worked with ABEC to define control parameters Single use bioreactor for virus expansion, study to support stability for extended hold time Clarification and Extraction retrofit Retrofit centrifuge to collect solids Two-step extraction, study to support no impact Modular depth filtration system optimized surface area with vendor 24
Downstream Optimization
Chromatography – Adapt to existing and new equipment, optimize for timing Changes to bed high, increased flow rates Study show no impact on performance Campaigning oversized buffer making and storage capacity Buffer stability at lab scale Sanitary maintenance protocol in production Increased surface area for DNA removal and TFF systems Study to support no impact at wider load ranges Integration of Single-Use Systems – End of Process Easy segregation from prior product contact surfaces Small scale unit operations, solutions and bulk storage Impact assessment (L&E) followed by study 25
• Target 30M doses
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses
• Yield Increase 10X
• Fast, 1-2 years
• Increased cadence
licensed process • Capacity to expand
Flublok Protein Yields at 600L Pilot Scale and 2000L Large Scale (normalized within serotype) Pilot Scale
Large Scale 1.6 1.3
1.0
0.9
1.0
1.0
1.0
1.0
1.0 0.9
H1/Cal
B/Mass
B /Brisbane
H3/Texas
B/Mass - Improved
rHA Antigen
Comparable purity, stability and all characterization profiles (data not shown)
• Target 30M doses
Next Generation Process
Large Scale Manuf.
• Yield Increase 10X • Increased cadence
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Demonstrate product comparability by Analytical Characterization Following ICH Q5E guidance • Rational Direct scale-up of licensed process
Limited number of process changes Good process characterization package for key process changes • Comparability is a protocol driven activity • Pre-defined acceptance criteria, Need to generate a base line for the specific antigens being used • Commercial and validation batches start at the same time, • Data was generated in parallel
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Mycoplasma/Spiroplasma Adventitious agents Infectious Baculovirus Potency Microbial enumeration Total Protein Appearance SRID/BCA Ratio Endotoxin Purity Identity rHA Size (SEC) pH Host Cell Protein Total DNA
Dynamic light scattering Flow microscopy SEC (UPLC) SDS-PAGE Trypsin Resistance RP-HPLC Hemagglutination ELISA Electron Microscopy DSF
• Four (4) weeks to GMP
• Anchor Point – 500L
production
• Efficient scale-up
• Universal Process • Ready to optimize for strain changes
• Yield Increase 10X • Cadence 1.75 batch/week
• 250K doses
Platform Technology
Product Launch
2nd Generation Process
Large Scale Manuf.
• Proof of Concept
• Consolidate as a Vaccine Manufacturer
• Target 2-5M doses,
• Formulation, delivery 29
• Target 30M doses
Next Generation Process
• Yield Increase 10X • Increased cadence
Clinical Studies • 2015 – Finalizing our Quadrivalent clinical trial for approval in 2016 • 2015 - Initiating our Panblok rH7 clinical trial – pandemic rHA • Pediatric clinical trial
Yield Improvements • Fed-batch process – increase volumetric productivity • Molecular biology approaches – Increase specific productivity
Stability and Formulation Improvements • Identified mechanism to stabilize the protein – addressing it at molecular level • Room temperature stability • New delivery systems - patch
Commercial Challenge • Different product presentations , expanded markets and collaborations
BHAG - 30M doses in 5 years!
M
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Target 30M doses
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
• Yield Increase 10X • Increased cadence
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Development of Flublok followed a carefully constructed strategy focused on the key challenges inherent to the product, Understanding likely challenges during commercial production and scale-up, are key to develop robust processes that accommodate the product’s growth cycle,
The success of Flublok demonstrates the maturity and adaptability of the BEVS technology as a production platform for challenging vaccines and biologics. Most importantly, Flublok provides
a new and powerful weapon against a one of the most important public health issues, and
the fastest and most effective response to a potential flu pandemic
The availability of Flublok to fight Influenza is a testament to the relentless dedication and enthusiasm of Protein Sciences employees, partners and supporters This work was supported by Federal (United States Government) funds from the Department of Health and Human Services, Office of the Assistant Secretary for Preparedness and Response, Biomedical Advanced Research and Development Authority, under contract number HHSO100200900106C to Protein Sciences Corporation. The content of this publication does not necessarily reflect the views or policies of the United States Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.
Influenza vaccine challenges and our strategy to overcome
Some background on influenza and influenza vaccines How we address influenza “constant change” through the product life cycle Development, Commercial production and Production expansion
Final thoughts and acknowledgement 2
Meriden, CT FDA licensed, multiproduct facilities 50L & 600L up to 500K doses
Pearl River, NY (formerly Pfizer) FDA licensed Flublok & Panblok 2,000L ~ 5M doses Gifu, Japan (UNIGEN) cGMP 2x21,000L >30M doses Expanding to 6x21,000L 3
Sf9 cells
SF+ cells
Sf9 cells cloned in 1983 from IPLB-Sf-21AE cells (isolated in 1970) Grown in fetal bovine serum Deposited at ATCC in 1985 by Texas A&M University (Cherry/Smith)
At PSC
Selective pressure in serum-free media in 1993 Unique phenotypic and genotypic properties Produce high titer baculovirus Patented Licensed for used in commercial production of USDA licensed veterinary vaccine and human products (US/Europe)
Autographa californica (Alfalfa Looper) Virus AcMNPV, E2 isolate AcMNPV polyhedra were isolated/cloned from a single-fieldcollected alfalfa looper larva Introduced restriction sites to facilitate generation and cloning of recombinant viruses
At PSC Infected our expresSF+ with parental, modified AcMNPV Harvest the virus rich supernatant, frozen, qualified, and designated Master BaculoVirus Bank (MVB)
DNA is isolated from MVB, linearized and recombined with transfer plasmid to form Working Virus Banks
Versatility
Multiproduct facility design
Speed
Single robust cell line for all products
Maximize capacity utilization Cloning in 21 days
Reliable scale-up
Current scale 2,000L, partners scale up to 21,000L
Low cost
High yields, low-cost proprietary media
Safety
BV highly specific to insect cells
GMP Production
Process optimization for new antigens High-density fermentation
No handling of dangerous virus
Concentrated DS
No containment required 6
Influenza vaccine challenges and our strategy to overcome Addressing “constant change” through the Product life cycle
7
Major surface glycoprotein, coat of the influenza virus, Antibodies against HA protect against influenza – the primary active ingredient in licensed vaccine
Changes in HA lead to annual epidemics requiring annual update of vaccine WHO /VRBPAC select the strains annually for the vaccine manufacturing 8
Typically we can expect at least one vaccine strain to change with each season FDA Selected Vaccine Strains by Season Season
H1N1 strain
H3N2 strain
B strain
2015-16
A/California/07
A/Switzerland
B/Phuket
2014-15
A/California/07
A/Texas
B/Massachusetts/2
2013-14
A/California/07
A/Texas
B/Massachusetts/2
2012-13
A/California/07
A/Victoria/361
B/Wisconsin/1
2011-12
A/California/07
A/Perth/16
B/Brisbane/60
2010-11
A/California/07
A/Perth/16
B/Brisbane/60
2009-10
A/Brisbane/59
A/Brisbane/10
B/Brisbane/60
2008-09
A/Brisbane/59
A/Brisbane/10
B/Florida/04
2007-08
A/Solomon Islands/03
A/Wisconsin/67
B/Malaysia/2506
2006-07
A/New Caledonia/20
A/Wisconsin/67
B/Malaysia/2506
Traditional Production • • •
Grow pathogen (in eggs) Inactivate pathogen Purify antigen (i.e., the active ingredient)
Challenges • Long production cycle • Affected by avian influenza outbreaks • Egg adaptation of virus • Egg allergies
Cell Culture Vaccines • Cell growth • MDCK, Vero, PER.C6 • Infection, virus generation • Inactivation / Splitting • Formulate vaccine
Challenges • High level of containment • Adaptation of virus to new substrate
Challenges overcome by recombinant HA vaccine manufacture
Engineer BV with gene of interest Hemagglutinin
BV highly specific to insect cells Powerful promoter generates high yields
Insect cells are cultured in fermentors
Infect cells with engineered virus
Incubate infection for ~48 - 72 hours
Purify protein to >90% Protein forms rosettes Formulate with PBS into vaccine
Flublok® 2013 Approval - Validation of the Platform 11
Trivalent Recombinant HA (rHA) Antigen Vaccine Produced in vitro via insect cell culture/baculovirus technology platform HA matches wild type human virus, no egg-drift 3X active ingredient compared to regular egg vaccine
Flublok rHA Antigens Highly purified, typical purity >90% Correct 3-D structure, biologically active Hemagglutination activity Induce protective immune responses HA antibodies/surrogate marker of efficacy Neutralizing antibodies
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 600L
• Universal Process • Ready to optimize for strain changes
• Yield Increase 10X • Cadence 1.75 batch/week • Formulation, delivery
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand 13
• Four (4) weeks to GMP production • Universal Process • Ready to optimize for strain changes
Challenges for an Influenza Vaccine
• Target 30M doses
Platform Technology
Next Generation Process
Seasonal Strains – Trivalent and quadrivalent formulation Drug Substance Changes - rHA protein changes constantly and the process must accommodate these changes Potency assay reagents - provided by CBER for the season, time critical Annual License Approval – Supplemental BLA submitted annually in time to release, pack and distribute product by early 3rd Q
• Yield Increase 10X
• Increased cadence
From a Process Development Perspective Robust, reproducible process – Universal process, flexible and adaptable Well understood and controlled – Deliberately designed operational spaces to allow optimization for antigen changes Routine, proceduralized approach - Complete optimization, if required in an extremely short time window
• Four (4) weeks to GMP production • Universal Process • Ready to optimize
2. infect
Disc Stack Centrifuge
Cell Pellet
for strain changes
Platform Technology
Depth • Target 30M doses Filtration
Next Generation Process
• Yield Increase 10X • Increased cadence
1. seed
Protein Production
Harvest
Extraction
Clarification
Purification Capture
Purify
DNA removal
Ultrafiltration/Formulation
• Four (4) weeks to GMP production • Universal Process • Ready to optimize for strain changes
Surveillance and HA Sequence Analysis
Cloning of Influenza HA Genes into Transfer Vector
• Data analysis : SH change, CDC, WHO, IFPMA, etc. • Year around HA sequences analysis • 1st option – Cloning from influenza virus • 2nd option – Site Directed Mutagenesis of previously generated HA genes • 3d option – Gene synthesis
Strain Selection
• Target 30M doses
Platform Technology
Next Generation Process
• Yield Increase 10X
• Increased cadence
3 Days
Transfer Vector Insect Cell Transfection and Plaque Purification
Baculovirus Bank Generation
• Co-transfection transfer vector with MVS baculovirus linear DNA • Plaque purification and screening
• Minimum passages during virus scale-up • Stable in liquid N2 • Quality tests ; PCR, Western, SDS-PAGE, etc.
21 Days
Working Virus Bank
• Four (4) weeks to GMP production • Universal Process • Ready to optimize for strain changes
Step
Isotype variability (H1, H3, B)
Cell Culture
upstream
Protein Production
Next Generation Process
Variable • Yield Increase 10X parameters • Target 30M doses
• Increased cadence
universal yes
Harvest
downstream
Seasonal strain variability
Platform Technology
Infrequent
Cell viability at harvest
no
Extraction
yes
Infrequent
pH; [detergent]
Clarification
yes
no
DF Filter type
IEX
yes
Sometimes
pH; [salt]; column type; [detergent]
HIC
yes
Sometimes
pH; [salt]; [detergent]
DNA Removal
Universal
Ultrafiltration Formulation
Universal
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Efficient but
Platform Technology
Framed by the design space for each unit operation • Target 30M doses
• Yield Increase 10X
Next Generation Process
Product Launch
limited supply
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years
• Fully contained within the ranges of the validated process • Focus on updates to specific set-points, key requirements Meet established process performance Maintain quality attributes and stability profile Optimize yield • Driven by known changes in the rHA protein and process knowledge • Sometimes, no changes are required specially for rHA of same lineage • Increased cadence
licensed process
• Capacity to expand
Proceduralized approach – SOP driven work • Rapid assessment with a qualified scale-down model or at production scale • Clearly defined acceptance criteria determines need for optimization • Changes are implementation following Quality System and do no require regulatory approval • Developed in collaboration with the FDA
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Efficient but
Platform Technology
Product Launch
limited supply
Large 2013 - FDA approves Flublok for the prevention of influenza Scale Manuf in adults 18 - 49 years old • Target 30M doses
Next Generation Process
• Yield Increase 10X
• Increased cadence
.
• Target 3-5M doses
• Fast, 1-2 years licensed process
“The Evolution, and Revolution, of Flu Vaccines”
• Capacity to expand
http://www.fda.gov/ForConsumers/ConsumerUpdates/ucm336267.htm
First recombinant influenza vaccine The pandemic solution Only pandemic vaccine that can be quickly manufactured and/or transferred and manufactured in other countries
2014 - Flublok approved for adults 18 and older 2015 - Large Scale Manufacturing approved for production of Flublok and Panblok 19
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Target 30M doses • Yield Increase 10X • Increased cadence
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
20
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Yield Increase 10X • Cadence 1.75 batch/week • Formulation, delivery
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand 21
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
CMO - Expedient, costly, restricted
Greenfield Facility: Perfect Fit, high cost, long time
Adapt an Existing Facility: Challenging, but affordable & flexible •
Relatively minor expenses Ready for capacity expansion, Attractive lease and government support
Moved-in Dec. 3rd 2012; Approved May 2015! 22
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Process Design
Readiness
~ 1 month BARDA Proposal
Preparation for Lease agreement
Match or “Fit” the Process to facility & equipment
Scope, quote and ready for equipment instrument purchase
Define high level design As-Is, Adapt, New Gap Assessment Characterization vs. Design
Risk Analysis Gap Closure Program
Focus: efficiency, cost and short lead times Raw materials & components plan media, resins, etc.
Start-Up & Confirmation 100 days! 1st batch March 26 2013 Match process parameters, yield, purity Optimize & complete documentation
Move to qualification & preparation for PV 23
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Upstream Process & Equipment Bacterial fermentation train retrofitted for cell culture Install oxygen service, new impellers, spargers, and DO control system Worked with ABEC to define control parameters Single use bioreactor for virus expansion, study to support stability for extended hold time Clarification and Extraction retrofit Retrofit centrifuge to collect solids Two-step extraction, study to support no impact Modular depth filtration system optimized surface area with vendor 24
Downstream Optimization
Chromatography – Adapt to existing and new equipment, optimize for timing Changes to bed high, increased flow rates Study show no impact on performance Campaigning oversized buffer making and storage capacity Buffer stability at lab scale Sanitary maintenance protocol in production Increased surface area for DNA removal and TFF systems Study to support no impact at wider load ranges Integration of Single-Use Systems – End of Process Easy segregation from prior product contact surfaces Small scale unit operations, solutions and bulk storage Impact assessment (L&E) followed by study 25
• Target 30M doses
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses
• Yield Increase 10X
• Fast, 1-2 years
• Increased cadence
licensed process • Capacity to expand
Flublok Protein Yields at 600L Pilot Scale and 2000L Large Scale (normalized within serotype) Pilot Scale
Large Scale 1.6 1.3
1.0
0.9
1.0
1.0
1.0
1.0
1.0 0.9
H1/Cal
B/Mass
B /Brisbane
H3/Texas
B/Mass - Improved
rHA Antigen
Comparable purity, stability and all characterization profiles (data not shown)
• Target 30M doses
Next Generation Process
Large Scale Manuf.
• Yield Increase 10X • Increased cadence
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Demonstrate product comparability by Analytical Characterization Following ICH Q5E guidance • Rational Direct scale-up of licensed process
Limited number of process changes Good process characterization package for key process changes • Comparability is a protocol driven activity • Pre-defined acceptance criteria, Need to generate a base line for the specific antigens being used • Commercial and validation batches start at the same time, • Data was generated in parallel
• Target 30M doses • Yield Increase 10X • Increased cadence
Next Generation Process
Large Scale Manuf.
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Mycoplasma/Spiroplasma Adventitious agents Infectious Baculovirus Potency Microbial enumeration Total Protein Appearance SRID/BCA Ratio Endotoxin Purity Identity rHA Size (SEC) pH Host Cell Protein Total DNA
Dynamic light scattering Flow microscopy SEC (UPLC) SDS-PAGE Trypsin Resistance RP-HPLC Hemagglutination ELISA Electron Microscopy DSF
• Four (4) weeks to GMP
• Anchor Point – 500L
production
• Efficient scale-up
• Universal Process • Ready to optimize for strain changes
• Yield Increase 10X • Cadence 1.75 batch/week
• 250K doses
Platform Technology
Product Launch
2nd Generation Process
Large Scale Manuf.
• Proof of Concept
• Consolidate as a Vaccine Manufacturer
• Target 2-5M doses,
• Formulation, delivery 29
• Target 30M doses
Next Generation Process
• Yield Increase 10X • Increased cadence
Clinical Studies • 2015 – Finalizing our Quadrivalent clinical trial for approval in 2016 • 2015 - Initiating our Panblok rH7 clinical trial – pandemic rHA • Pediatric clinical trial
Yield Improvements • Fed-batch process – increase volumetric productivity • Molecular biology approaches – Increase specific productivity
Stability and Formulation Improvements • Identified mechanism to stabilize the protein – addressing it at molecular level • Room temperature stability • New delivery systems - patch
Commercial Challenge • Different product presentations , expanded markets and collaborations
BHAG - 30M doses in 5 years!
M
• Four (4) weeks to GMP
• Proof of Concept
production
• Anchor Point – 500L
• Universal Process • Ready to optimize for strain changes
• Target 30M doses
• Efficient but
Platform Technology
Product Launch
Next Generation Process
Large Scale Manuf.
• Yield Increase 10X • Increased cadence
limited supply
• Target 3-5M doses • Fast, 1-2 years licensed process • Capacity to expand
Development of Flublok followed a carefully constructed strategy focused on the key challenges inherent to the product, Understanding likely challenges during commercial production and scale-up, are key to develop robust processes that accommodate the product’s growth cycle,
The success of Flublok demonstrates the maturity and adaptability of the BEVS technology as a production platform for challenging vaccines and biologics. Most importantly, Flublok provides
a new and powerful weapon against a one of the most important public health issues, and
the fastest and most effective response to a potential flu pandemic
The availability of Flublok to fight Influenza is a testament to the relentless dedication and enthusiasm of Protein Sciences employees, partners and supporters This work was supported by Federal (United States Government) funds from the Department of Health and Human Services, Office of the Assistant Secretary for Preparedness and Response, Biomedical Advanced Research and Development Authority, under contract number HHSO100200900106C to Protein Sciences Corporation. The content of this publication does not necessarily reflect the views or policies of the United States Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.
