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Principles, Practice and Guided Evolution of Biologics Manufacturing Platforms
David BeattieR&D Director, Biotech Process Solutions
EMD Millipore, a division of Merck KGaA
CMC Forum Europe 2011Barcelona
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What is a Platform?
“A method, equipment, procedure or work practice that may be applied across multiple products under development or manufacture.”
Examples:• An expression system Chinese Hamster Ovary (CHO) cells
• A screening system high throughput based on robotics
• An analytical method imaged capillary electrophoresis
• A drug product formulation citrate + sucrose + Tween
• A mode of cell culture perfusion culture
• A purification unit operation affinity chromatography
• A complete process comprising multiple operations Platform DSP
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What is a Platform?
“A method, equipment, procedure or work practice that may be applied across multiple products under development or manufacture.”
mAb AmAb BmAb CmAb DmAb E
A single company making a variety of related drugs
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What is a Platform?
“A method, equipment, procedure or work practice that may be applied across multiple products under development or manufacture.”
Flu AFlu BFlu CFlu DFlu E
Flu AFlu BFlu CFlu DFlu E
Flu AFlu BFlu CFlu DFlu E
An industry making a variety of related drugs
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What are the Benefits?
• Development effort and overall costs reduced
• Time to clinic and therefore to market reduced
• Process improvements leveraged over many products
• Economies of scale for equipment, components and raw materials
• Failure rates during GMP manufacturing reduced over time due to accumulated process experience
• Procedures for in-process and batch release testing become routine,implying risk of errors reduced
• Faster turnaround in multiproduct facilities
• Submissions of INDs/IMPDs during early stage development are expected to be facilitated more readily
• Platform processes are suitable for a modular validation approach, which in turn leads to reduced efforts and costs
• Overall benefit for industry, health care system and patient
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Definition of PlatformBaseline purification and analysis strategy for future mAb products
Standardized materials and methods
IPC testing matrix and methods with minimum specifications
Purpose of PlatformReduce the time required for DSP development activities
Minimize material demand lead times
Maximize the utilization of process improvements
Establish a standard process for benchmarking to industry expectations
Merck Serono DSP Platform Project
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Unit Op Identification
Unit operations selected from historical success and common practicesPolish
Chromatography
Protein AChromatography
Capture
Low pHhold
Anion ExchangeChromatography
Bulk DrugSubstanceFormulation
Nanofiltration
UF/DF
To achieve optimal versatility, the platform consists of a primary and alternativeapproach for each operation
Bulk DrugSubstanceFormulation
Flexibility!
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Platform Process Control
Standardized IPC testing matrix
Promote reliable data comparison
Establish expectations for impurity removal for each operation
Establish generic specifications for drug substance
Analytical Method
IPC Monitoring
BDS Specification
PROA
LOW pH
AEX
POL
VRF
UFDF
BDS
Conc. (rPA) X X X N/A
Conc. (A280nm) X X X X X X X Product Specific
Purity (SEC) X X X X X X X >95% Monomer
Purity (PAGE) X X X X X X X >95% Product Bands
Isotypes (ICE) X X X X Compare to RS
HCP (ELISA) X X X X X X X <50ppm
DNA (qPCR) X X X X X <10pg/mg
FLrPA (ELISA) X X X X <5ppm
Bioactivity X 60-140% of RS
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Case Study: Anti-CD19
The first application of the generic process
Primary chosen for Capture, Low pH and Virus Reduction FiltrationAlternative chosen for Polish for removal of aggregate to meet purity specificationAlternative chosen for UFDF based on yield, placed before VRF due to high NaCl
cHA
MabSelect Xtra
Low pHhold
SartobindQCharged
Membrane
Bulk DrugSubstance
Formulation
CPV
30KdRC
Process Recovery
Product Purity
Residual Aggregate Process Impurities
>65% >99.5% <0.5%All meet spec
or <LOQ
Repeated for other mAbs Success! (?)
FIMProcess
Development
cGMP API prod.
DP prod.
Pre-clin supply APIrelease
Tox material
Clone selected
Supportive stability studies
Formulation selection
DP release
cDNA available
Pre-MCB tested
Clone generation
Process check
Pre-clinical formulation
Analytical dev
Analytical check
regulatory documentation
MCB available
Stability studies
IND IMPD
Formulation check
Year 2Year 2
Aug OctSep NovJunApr JulMay FebDec MarJan Aug OctSepJunApr JulMay
Q2 Q3 Q4 Q1 Q2 Q3
Year 1Year 1
Feb MarJan
Q1
Nov Dec
Q4
Ph 1
Production
Benefits not realized in Isolation
FIMProcess
Development
cGMP API prod.
DP prod.
Pre-clin supply APIrelease
Tox material
Clone selected
Supportive stability studies
Formulation selection
DP release
cDNA available
Pre-MCB tested
Clone generation
Process check
Pre-clinical formulation
Analytical dev
Analytical check
regulatory documentation
MCB available
Stability studies
IND IMPD
Formulation check
Year 2Year 2
Aug OctSep NovJunApr JulMay FebDec MarJan Aug OctSepJunApr JulMay
Q2 Q3 Q4 Q1 Q2 Q3
Year 1Year 1
Feb MarJan
Q1
Nov Dec
Q4
Ph 1 Production
And Release
Cell Line
Generation
Analytical
Development
Formulation
Development
Benefits not realized in Isolation
30+ mosto FIM !
Lesson Learned: full benefit only from “Total Chain” Platform
Bulk DrugSubstanceFormulation
Purification Platform
Several sorting rounds
Cloning
Sorted cells growth
Staining and sorting highest fluorescing
cells
Stable pool
Several sorting rounds
CloningCloning
Sorted cells growthSorted cells growth
Staining and sorting highest fluorescing
cells
Staining and sorting highest fluorescing
cells
Staining and sorting highest fluorescing
cells
Stable pool
Cell Line Generation Platform
Analytical Platform
Formulation Platform
18 mosto FIM !
Benefits not realized in Isolation
Early Phase Development replaced by Platform Fit and Adaptation
Involve all the Stakeholders
Hydroxyapatite: calcium-phosphate crystal with a zero point charge at neutral pH Extremely sensitive to acidic conditions
Extremely powerful for polishing mAbs !
In non-Platform process, column degradation occurs after cycling
Visible loss of resin and bed cracking
At lab scale, up to 50 cycles run without issue
But, rapid and limiting increase in back-pressure only at production scale
Lesson Learned—manufacturing colleagues need a strong voice in setting Platform and non-Platform information can be relevant
Cycle 1 Cycle 20 Cycle 50
0.0
0.5
1.0
1.5
2.0
2.5
3.0
2 4 6 8 10 12 13 14 15 16 17 18 19 20 21
Batch No
Col
umn
inle
t pre
ssur
e (B
ar)
load
elution
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Using the Platform to create a Viral Clearance Knowledge Base
Development History and Prior Process Knowledge are core to QbDJust as we use accumulated performance knowledge from multiple
projects to design the Platform, we also look to accumulated virus clearance data to optimize the effectiveness
0
5
10
15
20
25
30
5 5.5 6 6.5 7 7.5 8 8.5 9
pH
Con
duct
ivity
(mS/
cm) MLV> 4
MLV<4MVM>4MVM<4SV40>4SV40<4Reo3>4Reo3<4
Clearance factors in function of the pH and the conductivityObjectives:Improve design space using viral clearance as response factorAmortize clearance data over many projectsReduce validation workload during early clinical phasesEfficient use of data accumulated on generic platform process(es)
Reconcile two apparently contradictory needs of process development: reducing cost and duration of development while simultaneously increasing process understanding
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Platform Evolution
“A platform process should not be regarded as a static, long-lasting procedure. In contrast, it should be reviewed within agreed life-cycle periods and undergo changes by controlled implementation of improvements and uptake of innovations.”
Moreover, “Processes of the Future” can and should be based on the learnings from the Platform
more Knowledge Management
mAb Chromatography Steps at 2 g/L
Protein A Virus Hold Cation Exchange
Hold Tank Anion Exchange
Hold Tank
Input12,0002 g/L
2,850 L8.2 g/L
3,280 L7.1 g/L
1,530 L13.7 g/L
3,060 L6.9 g/L
3,060 L6.9 g/L
Output2,850 L8.2 g/L
3,280 L7.1 g/L
1,530 L13.7 g/L
3,060 L6.9 g/L
3,060 L6.9 g/L
VirusUF/DF
Plant Design: 5,000L Tank 5,000L Tank 5,000L Tank
Platform Evolution: Process Compression
Protein A Virus Hold Cation Exchange
Hold Tank Anion Exchange
Hold Tank
Input12,0005 g/L
7,200 L8 g/L
8,300 L7 g/L
4,580 L11 g/L
9,160 L5.7g/L
9,160 L5.7g/L
Output7,200 L
8 g/L8,300 L
7 g/L4,580 L11 g/L
9,160 L5.7g/L
9,160 L5.7g/L
VirusUF/DF
Plant Design: 5,000L Tank 5,000L Tank 5,000L Tank
mAb Chromatography Steps at 5 g/L
Platform Evolution: Process Compression
What if you could remove intermediate hold tanks by direct loading of elution steps?
Protein A Direct loading of Pro A
elution and Virus Hold
on CEX media column
Cation Exchange Direct
loading of CIEX elution
on to membraneadsorber
Anion Exchange
Hold Tank
Input12,0005 g/L
7,200 L8 g/L
4,800 L10 g/L
4,800 L10 g/L
Output
7,200 L8 g/L
4,580 L11 g/L
4,800 L10 g/L
VirusUF/DF
Plant Design: 5,000L Tank
Platform Evolution: Process Compression
PoolHCP(ppm)
Leached Protein A
(ppm)
Aggregate (%)
Yield(%)
Feed (MAb spike/non-expressing CHO)
295,000 NA NA
Protein A Pool (ProSep Ultra Plus)
140 12 0.5 98
Cation Exchange Pool(Eshmuno S)
64 < 2* 0.3 83
Anion Exchange Pool(ChromaSorb)
< 3* < 2* 0.3 96
Overall Connected Process
< 3* < 2* 0.3 78
Overall Control Process (with intermediate holds)
< 3* < 2* 0.4 86
Performance Results from Direct Elution Connected Process
Platform Evolution: Process Compression
Comparable performance for Connected Process
Chromatography Step
Column Volumes (CV)
Bufferon-column
inactivation
Virus Conc.(Log)
on-column inactivation
BufferTraditional
Process
Virus Conc.(Log)
TraditionalProcess
CIEX EQ 450mM Sodium Acetate,
25mM NaCl, pH 5.4
50mM Sodium Acetate, 25mM
NaCl, pH 5.4
Load6
(load density = 50mg/mL)
Protein A Pool 6.5 Protein A Pool 6.5
CIEX Wash 350mM Sodium Acetate,
150mM NaCl,pH 3.5
≤ 2.450mM Sodium Acetate, 25mM
NaCl, pH 5.4
≤ 2.7
Low pH Hold
0.5(Total step time =
0.5hr, for on-column
inactivation)
50mM Sodium Acetate, 150mM NaCl,
pH 3.5≤ 2.8 Not performed N/A
CEX Elution 6*50mM Sodium Acetate,
w/NaCl,pH 5.4
≤ 2.250mM Sodium
Acetate, w/NaCl,pH 5.4
2.6
Post Elution 3EQ and EQ w / 1M NaCl
Pooled≤ 2.2
EQ and EQ w/ 1M NaCl Pooled
3.5
Platform Evolution: Process Compression
Virus Clearance Results from Direct Elution Connected Process
Below LOD for Wash, Hold, Elution and Strip
0
5
10
15
20
25
MAb A (1
1.4 g/L
)
MAb B (5
.2 g/L
)
MAb C Lot
A (18 g
/L)
MAb C Lot
B (18 g
/L)
MAb D Lot
A (26 g
/L)
MAb D Lot
A (13 g
/L)
MAb D Lot
B (15 g
/L)
MAb E Lo
t A (1
4 g/L)
MAb E Lo
t A (7
g/L)
MAb F (6
g/L)
MAb G (6
g/L)
MAb H (4
.5 g/L)
MAb I (4
.5 g/L
)
MAb K (1
0 g/L)
MAb L (1
0 g/L)
MAb M (6
g/L)
MAb N (2
g/L)
MAb O (2
g/L)
MAb Q (1
5 g/L)
MAb R (8
g/L)
MAb S (1
4 g/L)
MAb T (5
g/L)
MAb U (1
2 g/L)
MAb X (1
7.6 g/L
)
MAb Y (2
2.5 g/L
)
MAb Z (1
g/L)
Mas
s C
apac
ity (k
g/m
2)
Avg. 6.5 kg/m2 Mass Capacity over Protein concentrations 1-26 g/LClearly not robust !
Evolution: Improving Robustness
Evaluation of Virus Removal Fitration for multiple mAbs
0
2000
4000
6000
8000
10000
12000
14000
16000
MAb1MAb2MAb3MAb4MAb5MAb6MAb7MAb8MAb9MAb1
0MAb1
1MAb1
2MAb1
3MAb1
4
MAb
Mas
s th
roug
hput
(g/m
2)
Vpro alone
V-Pro with CEX shield filter
Evolution: Improving Robustness
Used Platform Knowledge to use a Cation Exchange shield filter to remove aggregates that were primary limiter of capacity
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Where are We Now?
• Development effort and overall costs reduced
• Time to clinic and therefore to market reduced
• Process improvements leveraged over many products
• Economies of scale for equipment, components and raw materials
• Failure rates during GMP manufacturing reduced over time due to accumulated process experience
• Procedures for in-process and batch release testing become routine,implying risk of errors reduced
• Faster turnaround in multiproduct facilities
• Submissions of INDs/IMPDs during early stage development are expected to be facilitated more readily
• Platform processes are suitable for a modular validation approach, which in turn leads to reduced efforts and costs
• Overall benefit for industry, health care system and patient