Pharma&Biotech
Flexible Bioreactor Facility Design from Pilot
to cGMP Manufacturing
IBC’s Flexible Facilities, April 2-4 2013, San Francisco
David
Valentine
MSAT) / Lonza, Slough UK
Elise-marie
Seng(Principle
/ , Basel Scientist,
/ 10. Mai 2012
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:



Global leader in microbial control and custom manufacturing:





45 major production and R&D sites
Employs over 10,000 people
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)
Slough, UK
Portsmouth, NH
Porriño, SP
200 – 2,000L Capacity
Tuas, Singapore
5,000 – 20,000L Capacity
10,000L Capacity
200 - 20,000L 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






Mammalian Biologics Centre of Excellence
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
800L STR
1000L SUB
2000L ALR
3 x DSP
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
2
1
Voice of the
Customer
Voice of the
Scientist
•CQA/TPP
•Process experience
•Yield requirements
•Regulatory expectations
•Translating VOC
•Experience (platform)
•Design and development
•Scientific solution
Processes
Operate ‘on
target minimum
variation’
3
4
Voice of the
Process
•Daily Process Control
•Process understanding –
technical/operational
•Process capability
Voice of the
GMP Plant
•NOR
•Equipment capability
•Asset age/replacement
•New technology
Development, Improvement and Control
of the Manufacturing System
dFMEA
•
•
•
•
•
Audit ‘as is’
process step
Go to Gemba
Actions
Parameter Name
Notes
Effects of failure
mode
Severity
Low
No impact expected
1
High
Decreases flux and
increases processing
time, could affect PQ
(increase aggs)
Low
No failure mode
Occurrence Detectability
RPN
Comments/recommendations for other studies
(any known edge of failures)
Parameter
Designation
Millipore Biomax or Pall 30KD Ultrafiltration
Cassettes
Cassettes
Membrane loading (g product/m2 )
1
1
1
Process characterisarion studies to be performed, no spec in
PPD currently
9
3
5
135
N/A
N/A
N/A
N/A
Membrane lifetime
High
May Impact yield and
cycle time
Low
5
1
1
5
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
Low
Not a failure mode
N/A
N/A
N/A
N/A
High
low impact
1
1
1
1
Membrane lifetime (storage)
Immediate Risk Mitigations
Short Term Projects
Long Term Projects
Investment Projects
New Technology Projects
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.
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
occurrence rating will be site dependent
Production Area Temperature
WFI Flush
Conductivity of permeate following WFI
flush
NWP (Normalized Water Permeability
Following it with equil buffer so severity is lowered.
Low
Failure
High
Not a failure mode
5
1
1
5
N/A
N/A
N/A
N/A
>80%
discard membranes when fail NWP
System Equilibration
Buffer Type
Low
System not fully
equilibrated
High
Not a failure mode
Low
3
1
1
3
N/A
N/A
N/A
N/A
System not fully
equilibrated
3
1
1
3
High
System not fully
equilibrated
3
1
1
3
Low
System not fully
equilibrated
3
1
1
3
High
System not fully
equilibrated
3
1
1
3
Equilibration buffer volume
System equilibration conductivity
System equilibration pH
Detailed Control
Plan
Process Flow Chart
Material
Preparation
Request Raw
Materials
Inputs
•
•
•
•
POD Kit
Disp Kit
Deviations
‘Just do it’ employee suggestions
Engineering work orders
Clinic cases - ‘Things stopped us
executing perfectly on time’
Generate New
Documents
Operation
Sub-Operation
Process
Harvest
Tank/Pallet
Truck
Flowmeter and equipment
Checks
Flowmeters
Check Raw
materials and
Documents
BR Prereq
Checks
Flowmeter
setup
Control Causes - Xs
Send sheet to tech services
Measure
Control Results - Ys
Is there a gap?
(yes/no)
Key Resp
Measurement/Evalu
ation Method
Sample
Frequency
Sampling Size
Analysis of Output
Action if out of Control
na
na
none
USP supervisor
na
every batch
na
none
reactive request to TS
nc
no
na
na
no
USP supervisor
na
every batch
na
none
reactive request to TS
nc
no
Process outputs
TS aknowledge reciept
Receipt of
spreadsheet
yes/no
email to fermentation
USP and MST check kit vs
excel request form and BR
and sign.
Check for differences
kit vs spreadsheet
yes
signed form
no control
no
na
MST
supervisor
Visual
every batch
100% Inspection
none
contact USP
nc
must match
MST and USP
Operator
visual
Every batch
100% Inspection
none
make comment supply
missing items
nc
no
USP operator
na
every batch
na
none
reactive collection of kit
nc
nc
nc
no
na
no control
no
na
na
no
Disp Operator
na
every batch
100% Inspection
none
reactive collection of kit
from disp
no control
no
na
na
no
USP operator
na
every batch
<100% Inspection
none
request missing items
from disp
Issues docs and get on plant
check for docs on
plant
yes
presence of docs on
plant
no
docs on plant
USP operator
visual
every batch
100% Inspection
none
reactive receipt of docs
nc
Colate documation at point
of use
no control
no
na
no
no
USP operator
na
every batch
na
none
move documents to point
to use
nc
Pod kits on plant and check
BR
check for kit on plant
yes
presence of kits on
plant
no
kit on plant
USP operator
visual
every batch
100% Inspection
none
reactive request of kits
nc
Dispensary kit on plant and
check BR
check for kit on plant
yes
presence of kits on
plant
no
kit on plant
USP operator
visual
every batch
100% Inspection
none
reactive request of kits
nc
nc
nc
Collect pallet truck
no control
no
na
no
no
USP operator
na
every batch
na
none
reactive collection of pallet
truck
Collect Stedim Tanks
no control
no
na
no
no
USP operator
na
every batch
na
none
reactive collection of
stedim
Ice collected and present in 70 Freezer
Check sample bags are
attached to fermenter
check for ice in
freezer
check for sample
bags
Check containers are
available for decon
no control
yes
Check integrity test unit is
available and working
no control
Select Apropriate flow
meters
presence of ice in
freezer
presence of sample
bags
yes
no
ice in freezer
USP operator
visual
every batch
100% Inspection
none
reactive collect ice
nc
no
Sample bags on reactor
USP operator
visual
every batch
100% Inspection
none
reactive attach samlpe
bags
nc
presence of containers
no
containers on plant
USP operator
na
every batch
na
none
reactive collection of
containers
nc
yes
Presence of working
tester on plant
no
tester on plant
USP operator
na
every batch
na
none
reactive collection of
tester
nc
no control
no
appropriate flow
meter selected
no
no
USP operator
na
every batch
na
none
unknowningly operating
outside of calibrated range
c
Label
date
in date
USP operator
visual
every batch
100% Inspection
none
obtain equipment within
expiry
nc
yes
Manual Clean of WFI flow
meter
Equipment
Checks
check expiry of raw materials
and clean/sanitised
equipment
pFMEA
Process or Product
Name:
Develop Daily
Control Plan
Location of Control
Criticality of Control (
C ). Is this a Key
Process Step
Specification
none
Collect (move to hall) tech
sevices harvest kit -1to2 days
prior to harvest
Disp deliver kit to the USP
ops 1 to 2 days pripr to
harvest.
USP ops official check of part
number against BR
Normal GMP
Operational
Experience
Measure
No control
No control
Process variables
No defined trigger to submit
request
Complete TS excel
spreadsheet from BR - Area
Supervisor
Pre-requisite activities
Daily Clinic
Review of
Performance
Failure
mode
UF/DF General Operating Variables
FMEA Date (Orig)
FMEA Review Process
Process Stage
Process Step or
Product Part
Check Avaiability of
Raws and
Equipment
Check tubing available
Potential Failure Mode
S
E
V
Potential Failure
Effects
O
C
C
Potential Cause(s) of
Failure
D
E
T
Current Controls
R
P
N
Actions Recommended
Area
Resp.
Check for measuring
cylinder
Check availabilty of correct
sized pump
Check for availabilty of
appropriate timer
check 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
Pick measuring cylinder
from 1st floor clean equip
prep room
Measuring cylinder not
available
Stop Process
6
Unable to autoclave tubing
Stop Process
6
Available tubing not suitable 1
Pump Calibration
Buffer not available
Return to facility
check required flow rate of
Not checked
titrant (down)
N/A
KanBan
6
36
None - as we have a second autoclave
KanBan procedure
6
36
None - procedures are acceptable
6
6
No tubing in MST
No tubing in stores
1
1
MST Operator
SAP
6
1
36
6
6
No tubing at supplier
1
SAP
6
36
None - as we have dual suppliers for
tubing
Different calibration
procedure
4
Available cylinders are not
suitable
4
Not aware of controls
6
96
Determine if a stocking policy is
available and set into KanBan system
MST
Colin Stretch
6
Buffer not made
6
Buffer make up
spreadsheet
6
216
Review effectiveness of new buffer
supply system and improve if required
DSP
Donal
O'Grady
Stop Process
6
Buffer not suitable
4
Buffer documentation
BRs
6
144
Review effectiveness of new buffer
supply system and improve if required
DSP
Donal
O'Grady
1
Operator distracted
1
In the BR
6
6
6
Operator distracted
1
In the BR
6
36
1
4
Pick the wrong measuring
cylinder
There are no cylinders
available in MST
1
Procedure in SOP for
flowrate
6
6
1
Not aware of controls
6
24
0
Incorrect flowate used
too low
Incorrect flowate used
too high
Unwrap tubing, using full
length Remove autoclave
label and add to BR
Attach tubing to buffer
None - procedures are acceptable
Stop Process
Modify calibration
procedure
Different calibration
procedure
Pick up buffers from buffer
prep area/storage
area/transfer hatch
1
Stop Process
Stop Process
Stop Process
0
Add tubing to pump head
0
Set pump speed
Prime tubing at maximum
speed
clamp off tubing when full
of buffer
0
0
0
Completion
Date
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.
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.
non-KPP
Planning for Quality and Control
Slough Site Vision - “To consistently deliver
value and excellence in the eyes of the
customer”
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
New Technology

Across the Manufacturing Process

Aligned to our Transformation plan
SUB Timeline: Track Record of
Market Launch vs. Lonza Interest
250L
SUB
MARKET
LAUNCH:
200L Wave
Latest TruBioDV
Dual Controller /
Bag re-design
1000L
SUB
50L
SUB
20L Wave
2000
2001
2002
2003
2004
2005
2006
2007
2008
2009
2010
2011
2012
2013
S20 Wave Bioreactor
LONZA
INTEREST:
S200 Wave Bioreactor
50L SUB (Pilot)
250L SUB (Pilot)
1000L SUB (Pilot)

LONZA has track record of evaluating
and implementing latest SUB
technology shortly after market launch
1000L SUB (GMP)
Dual
50/250/
500L
(Pilot)
Summary of SUB Evaluation
(2007)
Requirement
Comparable cell growth
Comparable productivity
Comparable metabolic profiles
(glucose, lactate, glutamine, ammonia)
Scale-up consistency
 mixing
 k La
 process control system integration
 between single-use and stainless steel
Comparable Product Quality
Result
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)
Temperature (°C)
pH control
Dissolved Oxygen
Tension (%)
Medium
Version 7 (v7)
Version 8 (v8)
36.5
Wide pH control boundary
Constant pH (narrow control
boundary)
pH shift (each set point with
narrow control boundary)
15
40
40
CD-CHO (LifeTech owned)
CD-CHO (LifeTech owned)
CM68 (Lonza owned)
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
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
Nutrient Feeds
SF41:
Continuous feed
Variable rate according to glucose
concentration
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
100
90
80
DOT (%)
70
60
50
40
30
20
10
0
0
24
48
72
96
120
144
168
192
216
240
264
288
Elapsed Time (h)
TruFluor (control)
ReUseable
312
336
360
Cell Culture Process Control
pH
7.30
7.20
7.10
pH
7.00
6.90
6.80
6.70
(no daily adjustments to single-use sensor)
6.60
0
24
48
72
96
120
144
168
192
216
240
Elapsed Time (h)
ReUsable
TruFluor
Bench pH
264
288
312
336
360
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 realtime 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, nonstandard 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