A Manufacturer’s Perspective on Innovations in Biomanufacturing
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Transcript of A Manufacturer’s Perspective on Innovations in Biomanufacturing
A Manufacturer’s Perspective on Innovations in
Biomanufacturing
Abhinav A. Shukla, Ph.D. Vice President
Process Development & Manufacturing KBI Biopharma, Durham NC
IBC’s Biopharmaceutical Development & Production Week, Huntington Beach, CA, 2013
Incremental vs. Disruptive Changes • Incremental changes improve upon existing
technology • Disruptive changes offer a new way of doing things • Both are important drivers of innovation
Gottschalk, Brorson & Shukla, Nature Biotechnology, 30(6), 489-491, 2012
Biopharmaceutical Innovation • ROI on biopharmaceutical products steadily
decreasing in the last 30 years • Approaching 8-10% cost of capital levels • Paradigm shifts occur most commonly when there is a
driver
What is driving change in the biopharmaceutical world?
• Increased demand for biopharmaceuticals • Both number of products and quantity produced • Biosimilars
• Reducing ROI on pharmaceutical investment • Increased competition
• Multiple drugs for the same target/indication • Expanding geographies for production
• Lower scales for commercial production • Personalized medicine • Higher cell culture titers
• Novel proteins and other biologics • PAT & QbD initiatives – maintaining high quality is an
ever present constraint
Increased demand for biopharmaceuticals Larger number of products • > 900 biopharmaceuticals in development for > 100
diseases • > $ 114 billion sales
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100 150 200 250 300 350
PhRMA Biotechnology Report, 2011
Increased demand for biopharmaceuticals Follow-on biologics
• All major markets have biosimilar legislation now • Comparability hurdles are being overcome • Large players are increasingly entering this segment
Decreasing ROI in pharmaceutical R&D investment • Average Internal Rate of Return on Investment (IRR)
7.5% vs. 12% in the late 1990’s • Heading towards cost of capital
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Deloitte Thomson Reuters, Fundamental Productivity Challenge in Pharma, 2011
Increased competition Multiple drugs for the same target/indication
Rheumatoid arthritis biologic drugs • Enbrel • Remicade • Humira • Orencia • Simponi • Cimzia • Actemra • Rituxan
Lower scales for commercial production Higher cell culture titers
• Increase in cell culture titers will drive downstream process innovation • Continuous separations & Non-chromatographic separations are growing
Lower scales for commercial production Personalized medicine • Smaller product volumes • Omics, metabolic profiling and systems biology will all grow personalized therapeutics • Cost of sequencing 1 Mbp of DNA now <$1 instead of $10,000 in 2001
QbD/PAT initiatives • Increased emphasis on product safety • Increased emphasis on process understanding • Increased use of statistics in the bioprocess space • Innovations in biosensors & “measure to control”
strategies
Innovations in the biopharmaceutical process & manufacturing space
• > 100 fold titer improvement in cell culture since rDNA technology began to be used for biopharmaceutical production
• Single-use manufacturing • Increasing dynamic binding capacity (DBC) of chromatographic media • Introduction of parvoviral grade filters & improvement of flux
properties of viral filters • Membrane chromatography • Continuous processing in bioprocessing • Cell free protein synthesis • Pseudo affinity separations on non-affinity stationary phases • Non-chromatographic separations • Scale-down process characterization & validation studies as a key
component of BLA/MAA filings
-Confidential-
Innovations in the biopharmaceutical process & manufacturing space
• > 100 fold titer improvement in cell culture since rDNA technology began to be used for biopharmaceutical production
• Single-use manufacturing • Increasing dynamic binding capacity (DBC) of chromatographic media • Introduction of parvoviral grade filters & improvement of flux
properties of viral filters • Membrane chromatography • Continuous processing in bioprocessing • Cell free protein synthesis • Pseudo affinity separations on non-affinity stationary phases • Non-chromatographic separations • Scale-down process characterization & validation studies as a key
component of BLA/MAA filings
• 1996: Introduction of the Wave bioreactor • 1998: Introduction of first membrane adsorbers • 2004: First 250 L disposable stirred-tank bioreactor • 2006: First 1,000 L disposable stirred-tank bioreactor • 2009: First 2,000 L disposable stirred-tank bioreactor • Advantages:
• Reduced risk of contamination • Reduced need for SIP (Steam in Place) • Reduced need for cleaning validation
• Now: • Several manufacturing facilities and production trains with end-to-end disposable technologies • Stainless steel facilities also make significant use of disposables for capacity extension and flexibility • Multiple vendors for each single-use unit operation
Disposable Manufacturing
Why are single-use 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.
-Confidential-
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
BPC
BPC
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)
-Confidential-
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
What is next for single-use systems? • Further expansion of scale (up to 5000L?) • Better systems for integrating unit operations
seamlessly • More vendors for downstream single-use technologies
(columns, UF/DF) • Improved biosensors for single-use systems • Improved standardization of systems
• Extractables studies from vendors • IQ/OQ documentation and system controls from vendors
-Confidential-
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.
How can existing downstream process steps be made more efficient? • 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 à disrupt impurity-product
interactions
Washes that disrupt protein-protein interactions
Conventional washes
Enhancing HCP clearance across Protein A • HCPs form a diverse set of impurities • HCP clearance is a key concern in biopharmaceutical
separation processes
• Conventional wisdom: use washes with a pH intermediate between load and elution solutions to wash the Protein A column post-loading
-Confidential-
Enhancing HCP clearance across Protein A
Washes can be developed to disengage HCPs from the product rather than disrupt product-Protein A ligand interactions
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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
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l Pro
tein
s (n
g/m
L)
Normalized Yield vs. normalized CHOP for a variety of washes on MAbSelect Protein A
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Yield normalized to control experiment
CH
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xper
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Direction ofdesired trend
Enhancing HCP clearance across Protein A • Use washes at high pH (pH > 7) to preserve Protein A –
mAb interactions • Include chaotropes in washes to disrupt HCP-mAb
interactions E v a lu a tio n o f in te rm e d ia te w a s h e s a t p H > 7 .0
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N o rm a liz e d yie ld % o f c o n tro l
No
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d C
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(% o
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Shukla, A. Hinckley, P. Host cell protein clearance during Protein A chromatography - development of an improved column wash step, Biotechnology Progress, 24, 1115-1121, 2008.
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
+
+ +
+ + Mixed
Mode
GE Healthcare, Capto MMC ligand
Ionic interactions
Hydrophobic interactions
Hydrophobic interactions
Ionic interactions
GE Healthcare, Capto Adhere ligand
Log k’ vs Log [NaCl]
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Log k'
Log [NaCl]
Lysozyme
pH 7.0
1M urea
5% ethylene glycol
50mM arginine
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pH 7.0
1M urea
5% ethylene glycol
50mM arginine
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Log k'
Log [NaCl]
Monoclonal an6body
pH 7.0
1M urea
5% ethylene glycol
50mM arginine
Wash development on mixed mode
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HCP (ppm
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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 • Pseudo-affinity separations by combining mixed mode interactions with highly selective mobile phase modulators
Process Analytical Technology • PAT: “a system for designing, analyzing and
controlling manufacturing through timely measurements (i.e. during processing) of critical quality and performance attributes of raw and in-process materials and processes, with the goal of ensuring final product quality” FDA Guidance
Hou, Y., Jiang, C., Shukla, A., Cramer, S. Improved Process Analytical Technology (PAT) for Protein A chromatography using predictive PCA tools, Biotechnology and Bioengineering, 108(1), 59-68, 2011.
-Confidential-
Innovations in Biopharmaceutical Process
Development & Manufacturing
“E pluribus unum”
Biochemical/Chemical Engineering/Biochemistry/Molecular Biology
Pharmaceutical Manufacturing
(continuous processing)
Statistics (QbD)
Materials Science (resins, membranes)