Industrialization of a stem cell process - How to identify the right Strategy - IBC Oct 2013

Copyright 2012 ATMI, Inc. All Rights Reserved. Copyright 2012 ATMI, Inc. All Rights Reserved.

Fabien Moncaubeig

Sr Application Specialist & Cell Culture Lab Associate

Workshop:

Industrialization of a stem cell process – How

to identify the right strategy?

October 21, 2013 – Bethesda, MD

IBC - Cell Therapy Bioprocessing

Copyright 2012 ATMI, Inc. All Rights Reserved.

ATMI Lifesciences where we come from

ATMI LIFESCIENCES

Is part of ATMI Inc, a global company market

leader in efficient process solutions for the semi-

conductor and lifesciences industry

ATMI support leading company in the GMP

manufacturing of vaccines, Mabs and protein

production since 10 years

ATMI is expert in clean manufacturing and supply

of integrated technologies for cell culture

ATMI leverage is core competency in cell culture

to develop innovative technologies and efficient

processes


Our know how

ATMI approach combines both Product

development , Process development & optimization

ATMI has two Applications & Development centers:

Fully equipped cell culture clean rooms.

45 experienced team including Engineers &

Bioengineers

Tech transfer to your facility or to your CMO

PD Services / Support implementation of our technologies

PD for New Bioreactor development

Application & Development Center

Brussels Minneapolis

Surface 500m² 100m²

Classification Class 10,000 (ISO

class 7)

Non-classified

Biosafety level BL-2 BL-2


What we want to offer to the cell therapy

industry

Complete solution for cell based product manufacturing

Offer a full solution for cell growth and harvest, based on the best technologies

availables

Leverage our expertise in cell culture and cell therapy process development to streamline industrialization of cell therapy products

Optimize and enable high volume manufacturing of cell based therapy

Develop the future solutions


Different scale-up and manufacturing challenges for

each segment Autologous

Low dose. cells per patient < 108

High dose. cells per patient > 108

Low # passages possible (Adult SC, chondrocytes, ...)

High # passages possible (hES, iPS,...)

Allogeneic

Patient to Patient variability

Multiple parallel batches

Logisitc

Donor reproducibility

Passage limitations

Targeted lot scale

Downstream and formulation


Your process = Your product

Has never been so true !

Parameters impacting product (cell) quality:

Shear stress 1

2D VS 3D growth 2

Growth Surface Material 3

Changing production platform may require

clinical trial bridging

1: Biotechnol Bioeng. 2007 Feb 15;96(3):584-95. - Effects of shear stress on 3-D human mesenchymal stem cell construct development in a perfusion bioreactor

system: Experiments and hydrodynamic modeling. - Zhao F, Chella R, Ma T.

2: Tissue Eng Part A. 2009 Jul;15(7):1763-73. doi: 10.1089/ten.tea.2008.0306. - Phenotype and gene expression of human mesenchymal stem cells in alginate

scaffolds. - Duggal S, Frønsdal KB, Szöke K, Shahdadfar A, Melvik JE, Brinchmann JE.

3: Acta Biomater. 2013 Feb 26. pii: S1742-7061(13)00105-0. doi: 10.1016/j.actbio.2013.02.035. - Why the dish makes a difference: Quantitative comparison of

polystyrene culture surfaces. - Zeiger AS, Hinton B, Van Vliet KJ.

Biological challenges to consider during

process development and scale-up

www.solohill.com

www.solohill.com


Mimic final scale conditions from the early

development steps

Evaluate impact of any process change

Increasing cell culture scale implies streamlining

and optimizing all related operations :

Media preparation

Fluid handling

Harvest

Concentration

Rinsing

Freezing

Filling

Increase scale implies indroducing and validating

new technologies

Technical challenges to consider during


Taking everything into consideration


Business challenges to consider for


Supporting product lifecycle

Accelerate time to market, reduce development time and ressources

Mitigating risk for market approval,

minimize changement

Supporting commercilization ramp up, offer flexible production

Offer an affordable product, implement a

cost effective manufacturing


21,000cm²

What are the technological options for

cell production

Scale/unit

Or

Or any other

suitable SUB

3D environment

Planar technologies 122,400cm² 18,000cm² 25,000cm²

http://www.google.com/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=hV5PuIZYk7HeDM&tbnid=DD0DEUUqAmpJbM:&ved=0CAUQjRw&url=http://www.youtube.com/watch?v=TZ3Ki3fxwS0&ei=aSlkUtXHHuPo2AWc5YHoDQ&bvm=bv.54934254,d.b2I&psig=AFQjCNFBQJUy9VnXw28m1-fpp5j1jv_9eQ&ust=1382382269457955


Technology choice is driven by batch size and # patients

50’s 10’s 100’s 500’s 5000’s >10000’s

Cells per lot

Patients per year

Allogeneics

Or Or any

other suitable

SUB

Future

technologies

Autologous


Technical and financial considerations

regarding Downstream process scale-up

Choice will be led by

Harvest volumes to process

Time constraint

Concentration factor and final targeted volume

For large volumes, combination of these technologies may be required

100L Unifuge 1700LATMI or Centrifuge bags <135ml

Fluidized bed

(i.e:Ksep)

Large scale

centrifuge

(i.e:Unifuge)

Small scale

centrifuge

(i.e:ATMI)

Tangential Flow

Filtration

Flow rate (ml/min) 400 – 9600 3000 500 20 - 12,500

Hold-up volume (ml) 100 - 1600 1700 135 100’s-1000’s

Disposable cost ($) 2000-2500*(Ksep

400) 3000 950 1500-3500$*

Process dvt effort + - - +++

* Jacob Pattasseril, Hemanthram Varadaraju, LyeTheng Lock, and Jon A. Rowley, “Downstream Technology Landscape for Large-Scale

Therapeutic Cell Processing”,BPI, March 2013,


Now, lets compare the pros and cons of 3

different scale-up approaches using,

Multilayers,

Xpansion,

Microcarriers


Technical considerations for a scale-up in

multilayer systems

Limited process development:

Multitray systems with headspace Almost no process dvt required

Compact multitray Additional optimization of fluid transfer and harvest protocol required

Scale-up strategy = Scale-out:

Max scale/unit = 25,440 cm² (Cell Factory 40)

Increase number of units

Linear increase of footprint (incubators)

Implementation of automated manipulator system can reduce FTE need

Cell observation can be challenging in the largest scale units (>36 trays)

Cm²/volumetric footprint


High footprint requirement

Requires high, early, investment in factory / CMO

High FTE cost per batch

Uneven FTE needs

High number of operations = high failure rate

Manufacturing and Business considerations for

a large scale multitray process

0

5

10

15

20

25

30

35

40

45

50

1

11

21

31

41

51

61

71

81

91

10

1

11

1

12

1

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1

14

1

15

1

16

1

171

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1

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1

25

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1

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1

281

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1

30

1

31

1

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1

33

1

FTE Hours required / day

Process B - Hyperstack 36 Process B – Xpansion 200

http://www.google.com/url?sa=i&source=images&cd=&cad=rja&docid=70_vTHTEZkYTgM&tbnid=Q64HWG9xQFFq1M:&ved=0CAgQjRwwADgO&url=http://www.lonza.com/go/literature/4318&ei=JeVjUtrpNInIyAHS3YFo&psig=AFQjCNGLfczxOCmP_51H439qXjs6AB_Y2Q&ust=1382364837920437


Technical considerations for a scale-up in

Xpansion

Linear scalability from 10 (6,120cm²) to 200 plates (122,400cm²)

Scale-up by maintaining linear speed constant

Controlled environment:

pH, DO, Temperature monitoring & control

Cell observation

Process development required to :

Optimize process parameters

Harvest protocol


Manufacturing and Business considerations for a large

scale Xpansion process

Minimize footprint compared to multilayer

4 Xpansion 200 fits in 1 incubator (42.5 x 34.5 x

87 inches)

Lower cost /batch than multitray systems

Lower FTE requirements

Higher Consumable cost/cm²

Batch Cost distribution

Hyperstack

CoGs

Xpansion



scale Xpansion process

Clinical trial phase 3

(300 patients)

FDA approval

Early commercialization

(1,000 – 25,000 patients)

Commercialization

(>50,000)

2014 2015 2016 2017 2018 2019

NEW Facility building

$ 3,732,885

Facility running under capacity Facility running full

capacity

CMO or current facility

NEW Facility building under

capacity

Facility running full

capacity

$ 3,463,142

CMO or current facility

Hy

pe

rsta

ck

36

X

pa

nsi

on

20

0

Xpansion enables to decrease investment and delays requirement for new facility


Technical considerations for a scale-up

with microcarriers 1/2 Very high scale lot size achievable:

6g cytodex1/L x 500L = 1,320m²

Developing a microcarrier process is challenging, requiring time and high expertise

level :

Process development challenges:

Identify scalable cell attachment and growth mixing conditions

Develop scalable harvest protocol (timings)

Maximize cells/ml (carrier density)

Develop a seeding train with bead to bead transfer

Microcarrier key selection criteria to consider:

Density, cm²/g, g/L achievable, coating

Impact on : mixing requirements, cells/ml

Bioreactor challenge:

Low shear while maintaining mixing capacity

Keep aggregates in suspension/movement

www.solohill.com


Technical considerations for a scale-up

with microcarriers 2/2



scale microcarrier-based process

Process development substantially longer and risky

Could delay phase III or early commercialization

Low footprint compared to any planar system

Limited Facility size requirement

Easy future capacity increase

Dramatically lower CoGs


Summary of technology scale capacity and

associated R&D efforts

Ana S. Simaria, Sally Hassan, Hemanthram Varadaraju, Jon Rowley,

Kim Warren, Philip Vanek, Suzanne S. Farid, Allogeneic cell therapy bioprocess economics and optimization: Single-use

cell expansion technologies, Biotechnologie & Bioengineering, released online on August 16th, 2013


A dual approach to cover short and long

term needs


Conclusion

No Universal answer

Multiple factors with specific weight depending on therapy and company status

R&D effort capacity

Targeted

batch size Short & long term

Patient number forecast

Clinical phase status

Process constraints

Cell production platform


Conclusion

Controlled

environment

(pH, DO, cell

observation)

Contaminat

ion risk

(#operatio

n/cm²)

Process

dvt

expertise

required

Process

dvt

duration

Cell quality

change risk

(initial

process in

TF)

Max scale

achievable

Required

Footprint

COGS

/cm²

Initial

Investment

required for

commercial

scale

Multitray system (10 or

40)

Compact multitray (36)

Xpansion

Microcarrier + SUB

Hollow fiber

Pros & cons summary table for large scale allogeneic therapies


Thank you


Case study Promethera introduction

May 2013 – Bio 2013 - ”Enabling cell therapy : scaling-up allogeneic stem cell manufacturing from multi-tray stacks to Xpansion™ multiplate bioreactor” – Eric Halioua –Fabien Moncaubeig


Case study Promethera process

comparaison


27

EMERGENCE

P0/P1

P3/P4: 4 CS-10

Phase I/II Process

Aseptic process

P4/P5: 2 x 5 CS10

P5R: 2 x 15 CS10

Centrifugation/Filling

Freezing

FORMULATION 3,4x109cells


Case study Promethera process

comparaison


28

EMERGENCE

P0/P1

P3/P4: 4 CS-10

P0/P1

P3/P4: 1 CS10

Phase I/II Process Phase II/III Process

P4/P5: 1 XP100

Aseptic process

P4/P5: 2 x 5 CS10

P5R: 2 x 15 CS10


Freezing

FORMULATION


Freezing

RECONSTITUTION

3,4x109cells

Closed process

22,1x109cells

P5R: 5 XP200

EMERGENCE


Case study Promethera – cost

comparaison


Industrialization of a stem cell process - How to identify the right Strategy - IBC Oct 2013

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Transcript of Industrialization of a stem cell process - How to identify the right Strategy - IBC Oct 2013