flexible bioreactor facility design from pilot to manufacturing
TRANSCRIPT
Pharma&Biotech
Elise-marie Seng / , Basel / 10. Mai 2012
Flexible Bioreactor Facility Design from Pilot
to cGMP Manufacturing IBC’s Flexible Facilities, April 2-4 2013, San Francisco
Pharma&Biotech
David Valentine (Principle Scientist, MSAT) / Lonza, Slough UK
Contents
Lonza Overview
Current Flexibility in Stainless Steel (example for context)
Slough Site Vision, Manufacturing Excellence and PfQC
Technology Transformation Plan and Site History of SUBs
Cell Culture Platform Processes (sets design requirements)
Case Study: A Fit-to-Process and Modular Single Use Bioreactor
System Installed in Pilot Plant
Next Steps and Summary
Lonza Overview
Life sciences driven company
Headquartered in Basel (Switzerland)
Sales of CHF 3.925 billion in 2012
Global operations: 45 major production and R&D sites
Employs over 10,000 people
Global leader in microbial control and custom manufacturing: Hygiene
Water treatment
Active pharmaceutical ingredients both chemical and biological
Cell therapy
Leading positions in product market niches: Endotoxin detection
Cell-based research products
Nutrition ingredients
Performance intermediates
Lonza Biologics - Mammalian Production
Capacity (>250,000L Globally)
Portsmouth, NH
Tuas, Singapore
Slough, UK
Porriño, SP
10,000L Capacity 5,000 – 20,000L Capacity
200 - 20,000L Capacity
200 – 2,000L Capacity
Stainless Steel Plant Context –
Flexible at 200L to 20kL GMP scale!
Singapore 2 (operational in early 2011) labeled as 20kL but has been
designed with an ability to harvest and purify:
2 x 200L (seed bioreactor as independent production system)
2 x 1kL (seed bioreactor as independent production system)
4 x 5kL (seed bioreactor as independent production system –up to 20g/L)
4 x 20kL (standard full volume of production bioreactor – up to 5g/L)
Stainless Steel Plant Context –
Flexible at the pilot scale!
Pilot mimics GMP Production across all sites
Evolved across history of Lonza mammalian bioreactor types
Airlift
Stirred tank
Adaptable primary recovery to fit scale and separation challenges
Established data set on process transfer and scale-up supports
interchangeability of bioreactor types
Slough, UK – Biologics Centre of
Excellence
Site Profile
Process Development
Technology Development
Analytical Services
Scale-up and transfer
Clinical and commercial GMP production
Manufacturing History
cGMP since 1983, expanded 1986 and 1994
(re-developed and upgraded 2012)
Multi-product
(antibodies and therapeutic proteins)
~30 products per year
10-15 new products per year
Pipeline to feed large scale plants
FDA, MHRA and PMDA approved
200L ALR 500L STR
2000L ALR 3 x DSP 1000L SUB
800L STR
Mammalian Biologics Centre of Excellence
Slough Vision
“To consistently deliver value and
excellence in the eyes of the
customer”
Where excellence is a highly desired state achieved through…
understanding customer needs through a strong collaborative
project management approach
leveraging our accumulated scientific and technological
experience and wisdom
a demonstrated track record of technology leadership and
scientific integrity
recognition from our employees that Slough is a great place to
work
continuously being driven toward flawless execution such that
our operational performance yields sustainable competitive
advantage
responsibility and accountability being readily accepted at all
levels
a systematic approach to managing our processes at cost levels
that make us competitive and profitable to sustain our business
Mammalian Biologics GMP Facility – Upgraded 2012
Product Lifecycle – The Systems View
Processes Operate ‘on
target minimum variation’
Voice of the Scientist
•Translating VOC •Experience (platform) •Design and development •Scientific solution
2
Voice of the Process
•Daily Process Control •Process understanding – technical/operational •Process capability
4
Voice of the GMP Plant
•NOR •Equipment capability •Asset age/replacement •New technology
3
Voice of the Customer
•CQA/TPP •Process experience •Yield requirements •Regulatory expectations
1
Development, Improvement and Control
of the Manufacturing System
Process or Product
Name:FMEA Date (Orig)
Process StageProcess Step or
Product PartPotential Failure Mode
Potential Failure
Effects
S
E
V
Potential Cause(s) of
Failure
O
C
C
Current Controls
D
E
T
R
P
N
Actions Recommended Area Resp.Completion
Date
Check Avaiability of
Raws and
Equipment
Check tubing available
Check for measuring
cylinder
Check availabilty of correct
sized pump
Check for availabilty of
appropriate timercheck for mixer base in
facility when eluting in to
Nalgene
check for levtech charging
unit in facility when eluting in
to Nalgene
Ensure Levtech unit is on
charge 1hr pre-use
Perform test of LevTech
Check availabilty of pH
down buffer
Leave facility
Pick tubing from Kanban Tubing not available Stop Process 6 Unable to autoclave tubing 1 KanBan 6 36 None - as we have a second autoclave
Stop Process 6 Available tubing not suitable 1 KanBan procedure 6 36 None - procedures are acceptable
Stop Process 6 No tubing in MST 1 MST Operator 6 36 None - procedures are acceptable
Stop Process 6 No tubing in stores 1 SAP 1 6
Stop Process 6 No tubing at supplier 1 SAP 6 36None - as we have dual suppliers for
tubing
Pick measuring cylinder
from 1st floor clean equip
prep room
Measuring cylinder not
available
Different calibration
procedure4
Available cylinders are not
suitable4 Not aware of controls 6 96
Determine if a stocking policy is
available and set into KanBan systemMST Colin Stretch
Modify calibration
procedure1
Pick the wrong measuring
cylinder1
Procedure in SOP for
flowrate6 6
Different calibration
procedure4
There are no cylinders
available in MST1 Not aware of controls 6 24
Pick up buffers from buffer
prep area/storage
area/transfer hatch
Buffer not available Stop Process 6 Buffer not made 6Buffer make up
spreadsheet6 216
Review effectiveness of new buffer
supply system and improve if requiredDSP
Donal
O'Grady
Stop Process 6 Buffer not suitable 4Buffer documentation
BRs6 144
Review effectiveness of new buffer
supply system and improve if requiredDSP
Donal
O'Grady
Return to facility 0
Pump Calibrationcheck required flow rate of
titrant (down)Not checked
Incorrect flowate used
too low1 Operator distracted 1 In the BR 6 6
N/AIncorrect flowate used
too high6 Operator distracted 1 In the BR 6 36
Unwrap tubing, using full
length Remove autoclave
label and add to BR
Attach tubing to buffer 0
Add tubing to pump head 0
Set pump speed 0
Prime tubing at maximum
speed0
clamp off tubing when full
of buffer0
FMEA Review Process
pFMEA
Operation Sub-Operation Control Causes - Xs Measure Control Results - Ys Measure Specification
Normal GMP
Operational
Experience
Is there a gap?
(yes/no)Key Resp
Measurement/Evalu
ation Method
Sample
FrequencySampling Size Analysis of Output Action if out of Control Location of Control
Criticality of Control (
C ). Is this a Key
Process Step
Material Preparation Process Process variables Process outputs
Pre-requisite activitiesNo defined trigger to submit
request
Complete TS excel
spreadsheet from BR - Area
Supervisor
No control none na na none USP supervisor na every batch na none reactive request to TS nc
Send sheet to tech services No control no na na no USP supervisor na every batch na none reactive request to TS nc
TS aknowledge recieptReceipt of
spreadsheetyes/no email to fermentation no no
MST
supervisorVisual every batch 100% Inspection none contact USP nc
USP and MST check kit vs
excel request form and BR
and sign.
Check for differences
kit vs spreadsheetyes signed form must match
MST and USP
Operatorvisual Every batch 100% Inspection none
make comment supply
missing itemsnc
Collect (move to hall) tech
sevices harvest kit -1to2 days
prior to harvest
no control no na na no USP operator na every batch na none reactive collection of kit nc
Disp deliver kit to the USP
ops 1 to 2 days pripr to
harvest.
no control no na na no Disp Operator na every batch 100% Inspection nonereactive collection of kit
from dispnc
USP ops official check of part
number against BRno control no na na no USP operator na every batch <100% Inspection none
request missing items
from dispnc
Issues docs and get on plant check for docs on
plantyes
presence of docs on
plantno docs on plant USP operator visual every batch 100% Inspection none reactive receipt of docs nc
Colate documation at point
of useno control no na no no USP operator na every batch na none
move documents to point
to usenc
Pod kits on plant and check
BRcheck for kit on plant yes
presence of kits on
plantno kit on plant USP operator visual every batch 100% Inspection none reactive request of kits nc
Dispensary kit on plant and
check BRcheck for kit on plant yes
presence of kits on
plantno kit on plant USP operator visual every batch 100% Inspection none reactive request of kits nc
Collect pallet truck no control no na no no USP operator na every batch na nonereactive collection of pallet
trucknc
Collect Stedim Tanks no control no na no no USP operator na every batch na nonereactive collection of
stedimnc
Ice collected and present in -
70 Freezer
check for ice in
freezeryes
presence of ice in
freezerno ice in freezer USP operator visual every batch 100% Inspection none reactive collect ice nc
Check sample bags are
attached to fermenter
check for sample
bagsyes
presence of sample
bagsno Sample bags on reactor USP operator visual every batch 100% Inspection none
reactive attach samlpe
bagsnc
Check containers are
available for decon no control yes presence of containers no containers on plant USP operator na every batch na none
reactive collection of
containersnc
Check integrity test unit is
available and workingno control yes
Presence of working
tester on plantno tester on plant USP operator na every batch na none
reactive collection of
testernc
Flowmeter and equipment
Checks
Select Apropriate flow
metersno control no
appropriate flow
meter selectedno no USP operator na every batch na none
unknowningly operating
outside of calibrated rangec
Manual Clean of WFI flow
meter
check expiry of raw materials
and clean/sanitised
equipment
Label date in date USP operator visual every batch 100% Inspection noneobtain equipment within
expirync
Process Flow Chart
Request Raw
Materials
POD Kit
Check Raw
materials and
Documents
Harvest
Tank/Pallet
Truck
FlowmetersFlowmeter
setup
BR Prereq
Checks
Disp Kit
Equipment
Checks
Detailed Control Plan
Parameter Name NotesFailure
mode
Effects of failure
modeSeverity Occurrence Detectability RPN
Comments/recommendations for other studies
(any known edge of failures)
Parameter
Designation
General Operating Variables
Low No impact expected 1 1 1 1
High
Decreases flux and
increases processing
time, could affect PQ
(increase aggs)
9 3 5 135
Low No failure mode N/A N/A N/A N/A
HighMay Impact yield and
cycle time5 1 1 5
Low Not a failure mode N/A N/A N/A N/A
High No impact expected 5 1 1 5
Low
Low risk to product,
may slow down
processing
3 3 5 45
High Presents product risk 3 3 5 45
WFI Flush
Low Not a failure mode N/A N/A N/A N/A
High low impact 1 1 1 1
Low Failure 5 1 1 5
High Not a failure mode N/A N/A N/A N/A
System Equilibration
Buffer Type
LowSystem not fully
equilibrated3 1 1 3
High Not a failure mode N/A N/A N/A N/A
LowSystem not fully
equilibrated3 1 1 3
HighSystem not fully
equilibrated3 1 1 3
LowSystem not fully
equilibrated3 1 1 3
HighSystem not fully
equilibrated3 1 1 3
Millipore Biomax or Pall 30KD Ultrafiltration
Cassettes
>80% discard membranes when fail NWP
UF/DF -
NWP (Normalized Water Permeability
Cassettes
System equilibration conductivity
System equilibration pH
Equilibration buffer volume
non-KPP
non-KPP
Priming Buffer non-KPP
1. Buffer is tested prior to release for production. Buffers not
meeting criteria for pH / conductivity cannot be released for
production use. 2. Following equilibration, processing cannot
continue unless system effluent meets the specified pH and
conductivity ranges. 3. Equlilibration of the system can be
repeated to ensure that the system conditions are acheived prior
to introducing the process stream to the system.
occurrence rating will be site dependent
Membrane loading (g product/m2 )
Process characterisarion studies to be performed, no spec in
PPD currently
Production Area Temperature
Membrane lifetime (storage)
Membrane lifetime
Cassettes given 12 month expiry upon first use, however NWP
test is completed pre and post each UF/DF.Cassettes stored in
0.1M naOH, supported by validated storage study
Membrane lifetime studies NWP test pre and post use. 12
month shelf. Expectation is that filter would be replaced if it
doesn't pass NWP. Severity would not impact product quality but
,ay impact process step performance.
Conductivity of permeate following WFI
flushFollowing it with equil buffer so severity is lowered.
1. Buffer is tested prior to release for production. Buffers not
meeting criteria for pH / conductivity cannot be released for
production use. 2. Following equilibration, processing cannot
continue unless system effluent meets the specified pH and
conductivity ranges. 3. Equlilibration of the system can be
repeated to ensure that the system conditions are acheived prior
to introducing the process stream to the system.
dFMEA
Actions • Immediate Risk Mitigations • Short Term Projects • Long Term Projects • Investment Projects • New Technology Projects
Inputs • Deviations • ‘Just do it’ employee suggestions • Engineering work orders • Clinic cases - ‘Things stopped us
executing perfectly on time’
Go to Gemba
Develop Daily
Control Plan
Daily Clinic
Review of
Performance
Generate New
Documents
Audit ‘as is’
process step
Planning for Quality and Control
We achieve this through…. •leveraging our accumulated scientific and technological
experience and wisdom
•having a profound understanding of cause and effect
relationships
•designing processes for manufacturability
•optimising processes at cost levels that protect
competitiveness and sustain profitability
•subjecting our processes to continuous improvement
•aligning the Technology Strategy with Process
Development and the Asset Lifecycle Management Plan
•Demonstrating a track record of scientific integrity and
technological innovation
Slough Site Vision - “To consistently deliver
value and excellence in the eyes of the
customer”
New Technology
Across the Manufacturing Process
Aligned to our Transformation plan
SUB Timeline: Track Record of
Market Launch vs. Lonza Interest
MARKET
LAUNCH:
LONZA
INTEREST:
2000 2001 2002 2003 2005 2006 2007 2008 2009 2010 2011 2012
S20 Wave Bioreactor
S200 Wave Bioreactor
50L SUB (Pilot)
250L SUB (Pilot)
1000L SUB (GMP)
Dual 50/250/ 500L (Pilot)
20L Wave
50L
SUB
1000L
SUB
Latest TruBioDV
Dual Controller /
Bag re-design
1000L SUB (Pilot)
200L Wave
250L
SUB
LONZA has track record of evaluating
and implementing latest SUB
technology shortly after market launch
2004 2013
Summary of SUB Evaluation
(2007)
Requirement Result
Comparable cell growth
Comparable productivity
Comparable metabolic profiles
(glucose, lactate, glutamine, ammonia)
Scale-up consistency
mixing
kLa
process control system integration
between single-use and stainless steel
Comparable Product Quality
Continuous Improvement of
The GS System™
Year Improvement
1992 The GS System™ Introduced
2001 pCON Vectors Introduced
2003 Launch of CHOK1SV Cell Line
2006 GS-CHO Protein-free Process (Version 6 media platform)
2006 pConPlus Vectors for Antibodies
2009 GS-CHO Chemically Defined Animal Component-free
(CDACF) Version 7 Media Platform
2009 Potelligent® CHOK1SV Cell Line Technology Launched
2011 GS-CHO CDACF Version 8 (v8) Media Platform
2012 GS Xceed™ Gene Expression System Introduced Globally
Summary of Processes
Process Version 6 (v6) Version 7 (v7) Version 8 (v8)
Temperature (°C) 36.5
pH control Wide pH control boundary Constant pH (narrow control
boundary)
pH shift (each set point with
narrow control boundary)
Dissolved Oxygen
Tension (%) 15 40 40
Medium CD-CHO (LifeTech owned) CD-CHO (LifeTech owned) CM68 (Lonza owned)
Nutrient Feeds
SF40:
Fixed continuous rate
Fixed volume
Duration: 4-5 days
SF50:
Continuous feed
Variable rate
Continued for
duration of culture
SF70:
Continuous feed
Variable rate
Continued for
duration of culture
SF41:
Continuous feed
Variable rate according to glucose
concentration
Glucose:
Continuous feed
Variable rate
Continued for
duration of culture
Glucose:
Continuous feed
Variable rate
Continued for
duration of culture
SF52, SF53 and SF54:
Bolus additions on
days 5, 8 and 11 after
inoculation
SF71, SF72 and SF54:
Bolus additions on
days 3, 6, 8 and 10 after
inoculation
Case Study:
Fit-to-Process Modular Bioreactor System
Outputs from -
Site Technology Transformation Plan
Planning for Quality Control (achieving ‘on target minimum variation’)
Equipment capability requirements (needs of latest platform processes)
Confidence in SUB performance as stepping-stone to larger scale (s/s)
Flexible bioreactor volume requirements to best meet customer needs
Compact and ergonomic operational space requirements
Energy use and environmental impact modelling
…have built a strategy that favours single-use systems at the Slough site
Fit-to-Process Controller
TruBioDV
Our choice of process control provider system (Finesse, TruBioDV), is cGMP compliant software also enables rapid user configuration without re-validation needs
Speed of implementation has been demonstrated in the pilot facility
Sets a precedence for facility installation time and a blueprint for
facility re-design
The software upgrade to TruBioDV version 4.5 + DeltaV version 11 now enables increased hardware capability as well as miniaturisation of control cart footprint
The flexible operation of TWO independent SUBs in parallel, chosen from set of THREE (50L, 250L or 500L shells) fits in a space smaller than ONE 500L stainless steel bioreactor
Fit-to-Process Design
Compact Equipment Arrangement
Design still allows easy access to points where operator interventions needed:
• Bag fitting
• Sensor connections
• Additive set-up
• Sampling and process monitoring
• Control interface
The Modular Pilot SUB System
at Installation – Dual Bioreactor, Central Controller
System configuration shows
250L (left) and 500L SUB (right)
inactive 50L SUB parked behind
Fit-to-Process Design
Feed Model
Standard platform process design for last 4 years requires:
Alkali for pH control
continuous+ variable feed 1 (glucose)
continuous+variable feed 2 (complex)
shot 1 (stable in acidic solution)
shot 2 (stable in alkali solution)
shot 3 (stable in neutral solution)
– these 3 shots are chemically incompatible, hence dedicated addition route
antifoam
7 inlets for feeds in total +1 for medium fill and inoc (always separated by time) required
This “8-inlet port” system is becoming a minimum design standard for Lonza bioreactors (stainless steel included)
Fit-to-Process Design
Feeding Automation
The increased capability includes 7 digital pumps for dedicated addition lines – no interventions once set up
‘2-click’ automation for addition of our platform process shots:
Alkali inhibited; correct shot order followed; pre-configured dose volume and time; monitoring of pH limits to pause if edge of acceptable range approached; totalising of quantities added and time outside alarms; ‘parallel charge’ option also available
Automation has enabled error reduction and improved consistency
Single-Use Sensors
Benefits
Single-use sensors fits the concept of single-use bioreactors
Overall cost of single-use sensor comparable to conventional
Facility turn-around time reduced by 2 days per run
Elimination of:
leaks
autoclaving
cleaning validation
drifting
mid-run sensors fails
Performance evaluation against conventional electrochemical sensors is currently in progress using the pilot SUB system
Cell Culture Process Control
Dissolved Oxygen
0
10
20
30
40
50
60
70
80
90
100
0 24 48 72 96 120 144 168 192 216 240 264 288 312 336 360
Elapsed Time (h)
DO
T (
%)
TruFluor (control) ReUseable
Cell Culture Process Control
pH
6.60
6.70
6.80
6.90
7.00
7.10
7.20
7.30
0 24 48 72 96 120 144 168 192 216 240 264 288 312 336 360
Elapsed Time (h)
pH
ReUsable TruFluor Bench pH
(no daily adjustments to single-use sensor)
Fit-to-Process Design:
More Controller Capability
Standard 6 digital mass flow valves allows variety of gassing strategies to be easily configurable
Triple DOT and pH inputs future-proof these key parameters (Triple is temporary - once optimal type chosen; revert back to dual)
pCO2 levels in this design of SUB have proved to be both beneficial (levels do not get too low), but also detrimental (levels can be higher, potentially supressing peak growth)
Jacketed SUBs and stand-alone heater+chiller thermocirculator units remove plant chilled water utility and provide equivalent thermal performance to stainless steel bioreactors
Single-use pressure sensor with an accuracy and sensitivity appropriate to a bioprocess bag bioreactor – provides improved early warning against bag pressure issues
Fit-to-Process Design:
More Controller Capability
Remote access via terminal server allows live viewing and controlling of system (from internal LAN, or laptop externally)
Allows roaming access to the same level of process control detail as an
operator on plant: Enables process experts to conveniently check system
details
Enables on-call operators to check system alarms prior to travelling to site,
and troubleshooting support from experts online
Desktop sharing with system administrators or vendor support enables real-
time connection, reducing the response time and need for travel for on-site
support
Generation of data in a compatible format for integration to a plant-wide network, common database, and linking to other ‘smart controller’ systems
SUB Pilot Upgrade Project Result:
A Synergy of Systems
The 3 elements of…
Cell Culture Platform Process Design
Process Control System Design
SUB Bag Design
…come together to set the benchmark for system capability
The project result has established a standard model that can be quickly replicated, offering flexibly from 25L to 1000L scale
Remote access and visibility of real-time and historical data enables process trending and control charting to ensure batch success
Batch success rate over first 6 months currently running at 100%
Next Pilot Plant Upgrade Project:
Install identical controllers on s/s bioreactors
Project underway to convert stainless steel 130L ALR and 400L STR to TruBioDV by October 2013
Upgraded process controllers will replace older, non-standard systems, and also provide capability to connect to a SUB shell, when s/s bioreactor not required enabling universal controller capability
This project will realise the concept of a standard process controller which envelops different types and scale of bioreactor – the bioreactor option will be the element that is easily swappable
Demonstrating success of new hybrid and flexible operations in the pilot plant justifies and drives change to replicate identical design in future GMP manufacturing bioreactor new-builds or retro-fits
Summary:
Where are we and where are we going
Completing pilot plant SUB and stainless steel upgrades to enable standardised controller and bioreactor system capability, then demonstrate success prior to replication in GMP manufacturing
Phased approach for new/retro-fitted equipment aligned to site asset replacement plan and business needs
Maintain hybrid and flexible mixture of stainless steel and single-use systems in Slough that are compatible with transferring processes to our largest plants globally
Learned that process complexity needs to be managed to ensure benefits of standardisation and process-fit to all plants / all scales are realised
Continue our Process for Quality Control as the system to steer operations to perform ‘on target with minimum variation’ and provide data to prioritise the implementation of specific new technologies that enable this
Thank you