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