orphan biopharmaceuticals & the cdmo
TRANSCRIPT
Orphan Biopharmaceuticals
& the CDMO
(Contract Development and Manufacturing Organization)
Abhinav A. Shukla, Ph.D.
Vice President, Process Development & Manufacturing
KBI Biopharma, Durham NC
Presented at: World Orphan Drug Congress, Washington DC, April 9-11, 2013
Why are orphan biopharmaceuticals unique? • Smaller material demand
• Fewer clinical batches reduced large scale manufacturing experience prior to BLA/MAA filing Flexible manufacturing at a smaller scale (< 2000L cell
culture volumes) needed (Single-Use Manufacturing Technologies) Increased focus on process knowledge from scale-down
experimentation (QbD)
• Limited ability to do clinical bridging studies • Process changes during clinical development are less
desirable since their clinical impact can often not be studied readily Getting the process right the first time (Building robustness
and scalability into the process right from the start)
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Biologics Commercialization
Pre-Clinical Phase I Phase II Phase III
Process Development Process
Characterization Process
Validation Process Monitoring
& Improvement
FIH Process • Deliver clinical process
quickly • Platform process • Clinical Supply
Submission & Approval
Lifecycle management
BLA Prep & PAI
Commercial Process • Deliver manufacturing process for
registrational trials and market • Design keeping large-scale manufacturing in
mind • Improve productivity, efficiency, robustness,
manufacturability, COGs • Analytical characterization and method
development
Process Characterization and Validation • Develop IPC strategy through understanding of process inputs and
outputs (design space) • Scale-down characterization and validation studies • Large-scale process validation to demonstrate process consistency • BLA preparation • Supporting documents for licensure inspections • Post-commercial process improvements (CI) • Post-commercial process monitoring
FIH process Commercial process
Gottschalk U., Brorson K., Shukla A. Nature Biotechnology, 30(6), 489-491, 2012
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Biologics Commercialization
Pre-Clinical Phase I Phase II Phase III
Process Development Process
Characterization Process
Validation Process Monitoring
& Improvement
FIH Process • Deliver clinical process
quickly • Platform process • Clinical Supply
Submission & Approval
Lifecycle management
BLA Prep & PAI
Commercial Process • Deliver manufacturing process for
registrational trials and market • Design keeping large-scale manufacturing in
mind • Improve productivity, efficiency, robustness,
manufacturability, COGs • Analytical characterization and method
development
Process Characterization and Validation • Develop IPC strategy through understanding of process inputs and
outputs (design space) • Scale-down characterization and validation studies • Large-scale process validation to demonstrate process consistency • BLA preparation • Supporting documents for licensure inspections • Post-commercial process improvements (CI) • Post-commercial process monitoring
FIH process Commercial process
A single development cycle Robust and complete process characterization package
Commercial manufacturing at smaller scales
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SINGLE-USE MANUFACTURING TECHNOLOGIES
Why are single-use manufacturing systems growing? • Lower capital and utility costs (up to 40% reduction*) • Increasing titers driving bioreactor scales smaller
• Single-use bioreactors now up to 2000L volume
• Increased universalization of biomanufacturing • Co-location of manufacturing with markets • Biosimilars (estimated $ 17 billion market by 2020) • Smaller market sizes for novel drugs in niche/personalized
applications • Market fragmentation making large single-product
manufacturing facilities redundant
• Single-use systems finding application in stainless steel facilities for enhanced operational flexibility
Laukel et al, BioProcess International, May 2011 Supplement, pp. 14-21.
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Media and Feed preparation utilizing disposable mixing, filtration and storage systems
Disposable shake flasks or disposable spinner flasks
MCB or WCB vial
Disposable expansion reactor
Disposable seed bioreactor
Disposable production bioreactor
Disposable fluid path centrifuge
Disposable depth filtration system
0,2 µm filter
Hold vessels (Bags)
Hold vessel (bag)
Disposable fluid path purification system
Disposable mixing tank
0,2 µm filter
Retentate
Permeate
PD
Disposable fluid path purification system
Disposable mixing tank
0,2 µm filter
BPC
Virus filter
BPC
0,2 µm filter
BPCBPC
Sterile bulk fill and sampling bags
Buffer preparation utilizing disposable mixing, filtration and storage systems
0,2 µm filter
Disposable fluid path UF/DF system
Aseptic connection
Hold vessel (bag)
Hold vessel (bag)
Hold vessel (bag)
Hold vessel (bag)
Hold vessel (bag)
Process Reproducibility
4 manufacturing runs in Single Use Bioreactors
Highly consistent process
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Scalability
•4 different scales •3L and 15L scales in non-disposable bioreactors
•Process performance with different working volumes is also reproducible
Single-use technologies in downstream processing • Centrifugation (kSep® Systems)
• Closed, continuous centrifuge with class VI product contact surfaces
• Counteraction of Centrifugal force and fluid flow force • Very low shear • Continuous operation • Reversal of flow direction empties the chamber • Up to 7.2 L/min
Single-use technologies in downstream processing • Depth filtration:
• Harvest depth filters have traditionally been single-use except for their holders
• Based on particle entrapment in a fibrous bed • Can be used as the primary cell separation step for smaller cell
culture harvest volumes • Millipore – POD® system • Pall - Stax® system • Sartorius – Sartoclear P ® • Cuno – Zeta Plus ®
Pall – Stax System
Millipore - POD
Single-use technologies in downstream processing • Chromatography
• Membrane adsorbers • Mustang® (Pall), Sartobind® (Sartorius), Chromasorb® (Millipore),
Adsept® (Natrix), • Q, S, HIC and salt-tolerant ion-exchange functionalities • Most widely used for trace impurity removal in a flow-through mode
(DNA, endotoxin, viral clearance) • Pre-packed chromatography columns
• ReadyToProcess (GE Healthcare), Opus (Repligen), GoPure (Life Technologies)
• Monoliths • CIM monoliths (BIA Separations), Uno monoliths (Biorad)
Up to 20 cm D available
Clinical and commercial manufacturing using single-use technologies • Smaller material demand drives reduced scale for
commercial manufacturing • Fidelity between clinical and commercial product
needed (ideally single facility that fits both needs) • Single-use manufacturing technologies reduce costs
and reduce risk of cross-contamination
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Shukla, A., Mostafa, S., Wilson, M., Lange, D. Vertical Integration of Disposables in Biopharmaceutical Drug Substance Manufacturing, Bioprocess International, 10(6), 34-47, 2012. Gottschalk, U., Shukla, A. Single-use disposable technologies for biopharmaceutical manufacturing, Trends in Biotechnology, 31(3), 147-154, 2013.
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QUALITY BY DESIGN (QBD)
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Quality by Design (QbD) • “Quality by design means designing and developing
manufacturing processes during the product development stage to consistently ensure a predefined quality at the end of the manufacturing process.” ICH Q10, FDA 2006
Process Design (Process Development)
Process Control Strategy Definition
Process Validation
Continued Process Verification
QbD
Critical Quality Attributes (CQAs)
Process Design Space
Linking CQAs to Clinical outcome
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Process design space Characterization Space
Control Space
Operating Range
Acceptable Range
Design Space
Process Parameters
Key Parameters
CPPs
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Integrative Approach Each step is influenced by the preceding step
Shake flask and seed bioreactor parameters may affect growth rate in the seed bioreactor.
Seed bioreactor and production bioreactor parameters may affect productivity and critical quality attributes.
Production bioreactor parameters may affect downstream steps.
Characterization studies are linked.
Vial Thaw
Shake Flasks Seed Bioreactor
Production Bioreactor
Downstream Steps
Biotechnology and Bioengineering, 106(6), 894-905, 2010.
Production Bioreactor
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Establishing A Process Control Strategy
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Application of Quality by Design (QbD): downstream resin lot variability
Biotechnology and Bioengineering, 107(6), 989-1001, 2010.
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Evolving expectations in Process Validation
• Q7A definition: “Process validation is the documented evidence that the process, operated within established parameters can perform effectively and reproducibly to produce an intermediate or API meeting its predetermined specifications and quality attributes” • FDA guidance, Jan 2011: “The collection and evaluation of data, from the process design stage through commercial production, which establishes scientific evidence that a process is capable of consistently delivering quality product” • Process validation is now viewed as a process that occurs throughout the lifecycle of a product
Process Design (Process Development)
Process Control Strategy Definition
Process Qualification
Continued Process Verification
Scale-Down Process Validation Studies
• Scale-down validation studies in addition to large-scale process validation (conformance lots)
• Probe extremes in the process and demonstrate them to be acceptable
• Examples • Reprocessing validation – combine hold times with process
conditions that create the greatest stress on the protein • Intermediate hold times – combine hold times and
demonstrate releasable drug substance • Viral clearance studies • Impurity clearance studies
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Validation of Host Cell Protein Clearance
Harvest
Column 1
Column 2
Column 3
Worst-case C1 eluate
Worst-case C2 eluate
Harvest
Column 1
Column 2
Column 3
Harvest
Column 1
Column 2
Column 3
Spiking Strategy • Some CHOP species in harvest
may not be encountered by C2 and C3 in Mfg
• LVR could be overstated for C2 and C3
Worst-case Strategy • CHOP species in eluate is relevant
to the next step • More accurate evaluation of LRV • Need process characterization to
identify worst-case condition
By-pass Strategy • HCP species in load are relevant to
that process step in case the previous step is by-passed (e.g. “resin bed channeling”)
• Represents most “challenged” scenario
Biotechnol. Progr., 24(3), 615 – 622, 2008
Worst-case harvest
Development Phase • Utilizing the right set of analytical tools for in-process
testing and release • Characterization assays are equally important • Utilizing a broad set of tools up front gives the best
chance of determining CQAs & linking them to the process
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Analytical Methods Portfolio • Protein Primary Structure
Peptide Sequencing via LC/MS/MS Amino Acid Analysis Peptide Mapping
• Biophysical Characterization CD, FTIR, DSC, DLS, fluorescence
spectroscopy
• Capillary and Slab Gel Electrophoresis CZE SDS-CGE cIEF and icIEF SDS-PAGE and IEF Western blot Microchip electrophoresis 2D gels and blots
• Glycan Analysis Oligosaccharide mapping Monosaccharide composition Sialic Acid Quantitation
• Process Residuals • ELISA (HCP, protein A etc.) • HPLC (antibiotics, IPTG, detergents, etc) • qPCR (DNA)
• HPLC • Size Exclusion (with MALLS) • Ion Exchange • Reverse Phase • Hydrophobic Interaction • Affinity
• Potency Assays • Binding Assays via ELISA, Biacore and
ForteBio • Cell Based Assays (e.g., proliferation,
cytokine release, etc.)
• Mass Spectrometry • Intact mass • Peptide mapping with LC/MS or
LC/MS/MS • Disulfide Mapping • Post translational modifications (e.g.,
oxidation, deamidation) • PEGylation site identification • Glycan Identification & site identification
• Particle measurements
• Visible & sub-visible particles
Comprehensive Analytics
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THE RIGHT SCIENCE FROM THE START
Designing more efficient HCP clearance into the downstream process • Most current chromatographic steps are designed to
remove impurities based on differential binding to the stationary phase surface
• Conventional wisdom: wash conditions are between binding and elution conditions
• Orthogonal approach disrupt impurity-product interactions
Washes that disrupt protein-protein interactions
Conventional washes
30
Enhancing HCP clearance across Protein A • HCPs form a diverse set of impurities • HCP clearance is a key concern in biopharmaceutical
separation processes
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Washes can be developed to disengage HCPs from the product rather than disrupt product-Protein A ligand interactions
96
116359243
34655
935491
05000
100001500020000250003000035000400004500050000
Null supernatant MAbSelecteluate (load =
nullsupernatant)
MAbSelecteluate (load =
null supernatant+ product)
Prosep A eluate(load = null
supernatant)
Prosep A eluate(load = null
supernatant +product)
Hos
t Cel
l Pro
tein
s (n
g/m
L)
Normalized Yield vs. normalized CHOP for a variety of washes on MAbSelect Protein A
0%
20%
40%
60%
80%
100%
120%
140%
0% 20% 40% 60% 80% 100% 120%
Yield normalized to control experiment
CH
OP
(ppm
) nor
mal
ized
to
cont
rol e
xper
imen
t
Direction ofdesired trend
Biotechnology Progress, 24, 1115-1121, 2008.
Do HCPs co-elute with the product or co-associate with the product?
Enhancing HCP clearance across Protein A
Enhancing HCP clearance across Protein A • Use washes at high pH (pH > 7) to preserve Protein A –
mAb interactions • Include selective modulators (moderate concentrations of
urea, ethylene glycol, salts, arginine) in washes to disrupt HCP-mAb interactions
Shukla, A., Hinckley, P. Host cell protein clearance during Protein A resin chromatography: development of an Improved wash step, Biotechnology Progress, 24, 1115-1121, 2008.
Evaluation of intermediate washes at pH > 7.0
0%
20%
40%
60%
80%
100%
120%
140%
0% 20% 40% 60% 80% 100% 120%
Normalized yield % of control
Norm
alize
d CHO
P (%
of co
ntro
l)
Mixed Mode Chromatography
• Takes advantage of more than one type of interaction • Can reduce process steps • Provides enhanced selectivity, “pseudo-affinity” • Several mixed mode resins have recently been developed with:
» Increased loading capacities » Higher ionic strength tolerance
+
+ +
+ + Mixe
d Mode
GE Healthcare, Capto MMC ligand
Ionic interactions
Hydrophobic interactions
Hydrophobic interactions
Ionic interactions
GE Healthcare, Capto Adhere ligand
Log k’ vs Log [NaCl]
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
2.60 2.80 3.00 3.20 3.40 3.60
Log
k'
Log [NaCl]
Lysozyme
pH 7.0
1M urea
5% ethylene glycol
50mM arginine
-0.40
-0.20
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
1.60
1.50 2.00 2.50
Log
k'
Log [NaCl]
RNase
pH 7.0
1M urea
5% ethylene glycol
50mM arginine
-0.20
0.00
0.20
0.40
0.60
0.80
1.00
1.20
1.40
2.10 2.30 2.50 2.70
Log
k'
Log [NaCl]
Monoclonal antibody
pH 7.0
1M urea
5% ethylene glycol
50mM arginine
Wash development on mixed mode
0
50
100
150
200
250
300
350
400
450
500
0.0% 20.0% 40.0% 60.0% 80.0% 100.0%
HCP
(ppm
)
Recovery
Capto MMC HCP Clearance 25mM Tris pH 7.0 (baseline)
25mM Tris pH 7.0, 5% ethylene glycol
25mM Tris pH 7.0, 50mM arginine
25mM Tris pH 7.0, 50mM NaSCN
25mM Tris pH 7.0, 1M urea
25mM Tris pH 7.0, 1M ammonium sulfate
25mM Tris pH 7.0, 0.1M NaCl
25mM Tris pH 7.0, 0.5M ammonium sulfate
25mM Tris pH 7.0, 0.1M NaCl, 1M urea
25mM Tris pH 7.0, 0.1M NaCl, 1M urea, 5% ethylene glycol
25mM Tris pH 7.0, 0.1M NaCl, 1M urea, 5% glycerol
• Selective wash strategies can eliminate one chromatographic step in non-mAb processes • Designing quality into the process
Designing processes with the end in mind • Having the right analytical methods and product quality
profile in mind from the start • Keeping issues that can be encountered in large-scale
manufacturing in mind from the beginning
Process yields & robustness
Titer & downstream yields Reproducibility
Column loading and buffer needs
Column loading drives costs!
Raw material selection Potential for variability
Supply assurance Compatible with cGMP
Process impact
Transfer ready processes
Processes that can be compatible with many scales and facilities
Conclusions • Orphan biopharmaceutical development needs
particular emphasis on • Developing a process with the end in mind (licensure filing)
to avoid multiple changes along the way • Manufacturing costs • Demonstrating process robustness without recourse to an
extensive manufacturing history
• A dedicated CDMO with the right knowledge and capabilities can help smooth the development pathway