Biodisposables

Utility and Technological Advances

Report 9215

June 2006

Authors: J. W. Terryberry and G. Thor

Research Director: Alfred Doig
Production Manager: Tracy Beaudoin

www.drugandmarket.com
One Research Drive, Suite 400A Westborough, MA 01581
Phone: +1 (508) 616-5500 Fax: +1 (508) 616-5544

About the Authors

J. W. Terryberry and G. Thor

Jeffery Terryberry has over a dozen peer-reviewed publications in Clinical Chemistry research.
Current areas of focus include orthobiologic drug development utilizing osteoinductive peptides,
monoclonal antibody production and immunogenicity, and the phytochemistry of novel herbal
medicines.
Gautam Thor received his PhD in Neurobiology. He has researched at UCSD in the field of
Neuroimmunology and is the author of numerous scientific and medical publications through
NeuroConsultants, and, is internationally recognized for his expertise in human diagnostics.
Biodisposables: Utility and Technological Advances

Copyright 2006 D&MD Publications

All rights reserved. No part of this document may be reproduced in any form or by any means,
electronic or mechanical, including recording, or by any information storage and retrieval
system, without written permission from the publisher.
First published in the United States by:

D&MD Publications
One Research Drive
Suite 400A
Westborough, MA 01581
Phone: +1 (508) 616-5500
Fax: +1 (508) 616-5544

http: / / www.drugandmarket.com

An informa publication.
ISBN: 1-57936-360-1
Published in the U.S.A.

Biodisposables

Table

of

Contents

Chapter 1: Executive Summary....................................................................

11

Chapter 2: Economics of Biodisposables: Efficient, Fast and Flexible

Product Development. ......................................................................................

21

Disposable Technology and Contract Manufacturing (CMOs). ................................22

Drivers for Single-Use Disposable Systems ..........................................................24
Disposable Bag TechnologyPacific GMP, AppTec, Stedim.....................................27
Market Implementation.......................................................................................28
Industrial Advantages of Disposable Components .............................................. 210
Disadvantages of Disposable Systems.................................................................. 211
Leachables and Extractables.............................................................................. 211
Elimination of Waste in the Biodisposable Context..................................................... 212

Return of Investment (ROI) Options................................................................... 212

A 100 1000L Pilot Facility Costs $10 - 50 million. ............................................. 213
Contract Manufacturer Xcellerex Case Study Advantages of Biodisposables............. 214
Process Comparison Models.............................................................................. 216
Model Estimation of Simulations. ........................................................................ 218
Monoclonal Antibody Production......................................................................... 220
Disposables Reduce Overall Cost of Manufacture. .............................................. 223
Conversion from Stainless Steel Equipment to Disposable Filtration.................... 227
Disposable FiltersFor Use in Chromatography.................................................. 230
Summary. ........................................................................................................ 233

Chapter 3: Disposable Bioreactors..............................................................

31

Scaling Up Logistics............................................................................................32
Comparison and Validation of Disposable Bioreactor Systems. ..............................33
Growing Cells in BagsWave Cellbags Characteristics........................................38
Factors that Amplify the Bioreactors Yield........................................................ 310
Wave Biotech Bioreactor Case Histories.............................................................. 311
CatchMabs Tsunami Bioreactor. ........................................................................ 316
Osmoteks LifeReactor. ..................................................................................... 319
Cellexus Biosystems. ........................................................................................ 320
Disposable Bioreactor Flasks, Bottles, and Other Containers.............................. 322
Integra Biosciences CELLine Biodisposable Flask .................................................. 323
FiberCell Systems FiberCell Hollow Fiber Bioreactor .............................................. 326
Hollow Fiber Bioreactor Applications:................................................................... 327
Synthecons Rotary Cell Culture System. .............................................................. 329
Fluorometrixs Cellstation .................................................................................. 330
Cellons RollerCell............................................................................................. 331
Integra Biosciences Cellroll................................................................................ 332

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Biodisposables

Cornings CellCube. ......................................................................................... 332

Multi Laminate Plastic Bags ............................................................................. 334
Stedim. .......................................................................................................... 334

Chapter 4: Downstream Processing and Separation Sciences...............

41

Disposable Downstream Processing ....................................................................41

Faster Processing..............................................................................................41
Cleaning Ease ...................................................................................................42
Disposable Filters...............................................................................................42
Disposable Mixing, Filling and SamplingHyNetic, Alfa Laval Biokinetic,
Millipore, LevTech, Acerta, Nova Aseptic. ............................................................44
Sartorius..........................................................................................................44
LevTech............................................................................................................44
Millipore...........................................................................................................45
NovAseptic . .....................................................................................................47
Sampling Systems.............................................................................................47
Disposable Sampling Pump .................................................................................48

Sterile Connectors and Clean Room Technology....................................................49

Steam-in-place (SIP) Connector........................................................................... 411
Fully Assembled, Pre-sterilized Filtration Systems. ................................................. 412
Millipores Mobius Disposable Systems . .............................................................. 414
Separation Sciences: Filtration and Chromatography.......................................... 416
Disposable Filter Systems.................................................................................. 416
Disposable Filtration Market Statistics................................................................. 417
Filtration Technology......................................................................................... 418
Enhancing the Efficiency of Disposable Filters........................................................ 419
Sterile FiltrationSartorius, Millipore, Pall Corp................................................ 421
Millipore Sterile Filtration................................................................................... 422
Sartorius........................................................................................................ 422
Pall Corporation.. ............................................................................................ 424

Disposable Filter Capsules and Chromatography Columns................................... 425

Membrane Adsorption Chromatography .............................................................. 427
Membrane Chromatography and Viral Clearance................................................... 433
Affinity Chromatography and Antibody Purification............................................. 434
Inline Filters for Disposable Bioreactors............................................................. 436
Recovery of Cell Products through Chromatography........................................... 437
Cross/Tangential Flow Filters in Cell Processing and Diafiltration........................ 440
Cross Flow Filtration......................................................................................... 440
Sartorius BBI Systems Inc................................................................................. 441
CellFlo Applications.......................................................................................... 441
Cross Flow Process Modes for Cell Processing........................................................ 442

Tangential Flow Filtration TFF from SciLogic.......................................................... 445

Blood and Serum Fractionation.......................................................................... 446
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Biodisposables

Blood Fractionation........................................................................................... 446

Serum Fractionation......................................................................................... 447
Biosep Accoustic TechnologyApplikon. ............................................................. 449

Chapter 5: BiodisposablesFacility Design and

Future Developments........................................................................................

51

Facility Design Capabilities with Disposables. .......................................................51

Multi-Component Systems...................................................................................55
Flexel 3D Mixing System with Temperature Control..................................................55
The Single-Use Rapid Aseptic Fluid Transfer (RAFT) System.......................................55
Kleenpak Single-Use Aseptic Connectors................................................................55
Celsius-Pak. ......................................................................................................55
Functional and Environmental Constraints. ..........................................................57
Optimizing Facility Design....................................................................................58
Suppliers. .........................................................................................................58
Biocompatibility..................................................................................................59
Production Volume. ............................................................................................59
Disposal. ..........................................................................................................59
Disposable Design Optimization. ......................................................................... 510
Regulatory Considerations for Disposable Component Validation. ........................ 512
Sample Regulatory Requirements. ...................................................................... 513
Future Outlook of Disposables........................................................................... 516
Disposable Product Innovations. ......................................................................... 518
Market Projections........................................................................................... 519
Bags, Bioreactors, Filters, etc.. ......................................................................... 519
Disposable Biosensors...................................................................................... 524

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Biodisposables

Table

of

Exhibits

Exhibit 2.1

Strategic Outlook for Biodisposables.......................................................... 21

Exhibit 2.2

Disposable Components and Suppliers for Manufacturing of Biologics .

and Pharmaceuticals.............................................................................. 22

Exhibit 2.3

Hierarchical Representation of Biomanufacturing Tasks ............................... 24

Exhibit 2.4

Reasons for Implementing Biodisposables.................................................214

Exhibit 2.5

Costs of a Model Monoclonal Antibody Plant..............................................220

Exhibit 2.6

Cost of Goods Cap-Ex, Materials, and Labor in a Disposable Module Plant .....221

Exhibit 2.7

COGS from 100 L Plant.........................................................................221

Exhibit 2.8

COGS from 1,000L Plant.......................................................................222

Exhibit 2.9

COGS from 10,000 L Plant....................................................................222

Exhibit 2.10 Analysis Reported by the FDA CBER on Cost Manufacturing.........................223

Exhibit 2.11 Capital Required and Cost Savings of Disposable Model...............................223
Exhibit 2.12 Cost Analysis of Reusable Fermenters vs. Disposable Bioreactors.................225
Exhibit 2.13 Input and Output Parameters of an Analysis Tool........................................225
Exhibit 2.14 Disposable Technology and Conventional Pilot Plant COG Analyses ................227
Exhibit 2.15 Comparison of Equipment and Consumables in the Model Study. ..................228
Exhibit 2.16 Annual (COG/g) on a Cost Category Basis for Pilot Plant, .
Disposables-based (DISP) Pilot Plant, and Hybrid (HYB) Pilot Plant.................229
Exhibit 2.17 Comparison of Set-up Times and Cost of Single-use Cartridge .
Filter and Stainless Steel Housing............................................................232
Exhibit 2.18 Comparison of Process Time and Cost for Traditional Column and .
Membrane Chromatography Capsule........................................................232
Exhibit 3.1

Types of Disposable Bioreactors .............................................................. 34

Exhibit 3.2

Principle of the Wave Bioreactor.............................................................. 37

Exhibit 3.3

Cellbag Connectors and Biocompatibility Testing.......................................... 38

Exhibit 3.4

Research, Pilot, and Production Scale Wave Bioreactors. ............................ 39

Exhibit 3.5

Comparison of Daily Recombinant Protein Yields (mg/L) for Three Wave

Bioreactors..........................................................................................310

Exhibit 3.6

Monoclonal Antibody Yields for System20/50EH. ......................................311

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Biodisposables

Exhibit 3.7

Carriers Used to Cultivate Attachment-dependent Cells in the .

Wave Bioreactor...................................................................................312

Exhibit 3.8

Adenovirus Production and Recovery in HEK293 Cells using a .

Wave Bioreactor with Cytodex-3 Micro Carriers ........................................313

Exhibit 3.9

Use of Floating Perfusion Filter with Perfusion Controller in .

Wave Bioreactor...................................................................................314

Exhibit 3.10 Comparison of Batch and Perfusion Modes for MAb Production in the .
Wave Bioreactor (System 20/50EH).......................................................315
Exhibit 3.11 Comparisons of Batch and Perfusion MAb Production with 2L Cellbag...........315
Exhibit 3.12 Tsunami Multiple Wave Agitated Culture System. .......................................316
Exhibit 3.13 Excellerex Disposable Bioreactors............................................................317
Exhibit 3.14 AppliFlex Disposable Cell Culture Bag System.............................................319
Exhibit 3.15 LifeReactor Plant Cell Bag Culture System. ...............................................320
Exhibit 3.16 Cellexus Biosystems Optimized Cell Growing Chamber.................................321
Exhibit 3.17 Bags are more Stable than Bottles..........................................................322
Exhibit 3.18 Integra CELLine Disposable Bioreactor Flask..............................................323
Exhibit 3.19 Cost and Yield for Monoclonal Production in CELLine and Other Systems. ......324
Exhibit 3.20 Comparisons of CellLine, Roller Bottle Grown, and Bioreactor. .....................325
Exhibit 3.21 FiberCell Module and Flowpath Assembly with Medium Reservoir Bottle.........328
Exhibit 3.22 Rotary Perfusion RCCS-D and -4D Disposable Bioreactors...........................329
Exhibit 3.23 Cellstation High-throughput Bioreactor with Disposable Culture Vessels. ........331
Exhibit 3.24 RollerCell40 Automated Bottle System.....................................................332
Exhibit 3.25 Cornings CellCube System....................................................................333
Exhibit 3.26 Growth of Cells under Different Seeding Densities in the CellCube System......334
Exhibit 3.27 Flexel 3D Bag Bioreactor Harvesting System.............................................336
Exhibit 3.28 Composition of Flexel 3D Bags ...............................................................337
Exhibit 3.29 Xcellerex Disposable Bags and Mixing Systems..........................................338
Exhibit 4.1

Downstream Processing Applications for Disposables.................................. 43

Exhibit 4.2

Disposable Fluid Paths: Final Fill and Finish ................................................ 46

Exhibit 4.3

Acerta 4 Head Filler .............................................................................. 47


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Biodisposables

Exhibit 4.4

NovAseptic Sampling System with 9-Port NovaSeptum Connector.................. 48

Exhibit 4.5

Palls Kleenpak Sterile Disposable Connector System. .................................410

Exhibit 4.6

Millipores Lynx S2S Sliding Gate Sterile Connector...................................410

Exhibit 4.7

Ready to Run SIP Connected System: Sterilization of the .

Connection Between Millipores Lynx ST and SS Tank Interface. ....................411

Exhibit 4.8

NovAseptic SIP Connected System with Welded Connectors and .

Disposable Sampling .............................................................................412

Exhibit 4.9

Matching Downstream Processing to Disposable Product............................415

Exhibit 4.10 Disposables in Separation Sciences. ........................................................417

Exhibit 4.11 Applications of Sterile Filtration. ..............................................................422
Exhibit 4.12 Applications of CUNOs LifeAssure Filters..................................................423
Exhibit 4.13 Disposable Manifolded Filtration System with SupraClean Depth Filters..........424
Exhibit 4.14 Phases of Biomanufacturing Utilizing Disposable Filters and .
Chromatography Units...........................................................................426
Exhibit 4.15 Typical Production Scale Enzyme Purification with Multiple Filtration Steps......427
Exhibit 4.16 Sartorius SingleSep Disposable Multiuse (Autoclavable) Capsules................432
Exhibit 4.17 Virus Elimination from Monoclonal Antibody Preparation (Eluate from Affinity
Column) Prior to Sterile Filtration and Fill: High LRV and MAb Recovery.........434
Exhibit 4.18 Antibody Downstream Processing............................................................436
Exhibit 4.19 Inline Filters for Disposable Bioreactors....................................................437
Exhibit 4.20 Online Affinity Purification System. ...........................................................439
Exhibit 4.21 CUNOs Bioassure PES Filters for Cell and Immunoglobulin Recovery. ............441
Exhibit 4.22 CFF Process Modes Affecting Cell Yield....................................................443
Exhibit 4.23 Cross Flow Hollow Fiber Membrane System for Aseptic Cell Processing.........444
Exhibit 4.24 Purification of Biopharmaceuticals Using Amersham .
Biosciences CFF Columns.......................................................................445
Exhibit 4.25 SciLogs Automated Bioprocessing System with Disposable TFF Filtration.......446
Exhibit 4.26 Disposable CUNO Filter Types Used In Multilevel Blood Fractionation. ............447
Exhibit 4.27 Comparison of Single-use and Stainless Steel BSA Purification Systems. ........449
Exhibit 4.28 BioSep Cell Retention System..................................................................450
Exhibit 5.1

Single-use Technology Implementation Approach. ........................................ 53

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Exhibit 5.2

Layout of Facility Implementing Disposables with Functional and .

Environmental Classifications. .................................................................. 57

Exhibit 5.3

Disposable Design Optimization ..............................................................510

Exhibit 5.4

Validation Testing Methods and Standards. ...............................................516

Exhibit 5.5

Automated Remote FlexFactory Operation.................................................523

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Biodisposables

Chapter 1: Executive Summary

The current economic and regulatory realities, highlighted by the imminent establishment
of drug pricing controls and tightening regulatory and quality standards, indicates the added
pressures that are emerging for pharmaceutical companies to restrategize their overall approach.
There is a trend toward fewer blockbuster drugs as patient populations become smaller, and the
associated histories and genetic make up become stratified as personalized medicine begins to
come into its own. Biodisposable manufacturers responses to these trends is to develop fully
integrated, turnkey manufacturingproduction line that combines single-use components with
modular software; a disposable stir-tank and mixing system; and disposable bioreactors, along
with single-use membranes, filters, bags, tubing, and connectors.
Several commercial scale, process steps have biodisposables at numerous points in the pilot
scale as they are well suited for a multi-product clinical manufacturing setting with frequent
product change-overs. Single-use technologies are not only available for an increasing range of
applications, but they are also expanding from stand-alone devices to multi-component systems.
With this growing trend comes a greater number of benefits and implementation considerations
from pilot scale modules to complete production facilities.
Advantages of utilizing biodisposables in pilot and commercial scale operations involve reduced
downtime for cleaning, sterilization, and process engineering, as well as improved equipment
lead time, and process utility requirements such as validation, quality and regulatory compliance.
Biodisposables are well established in the industry, and there is an increasing trend toward
their use because they contribute to ensure the production of a safe, therapeutic product.
Biodisposables decrease capital costs, cost of goods, and labor, improve operability, flexibility,
and optimize utilization of space. Furthermore, components that integrate biodisposables with
stainless steel allow hybrid plant operation.
The average time required to construct a biotech facility is about five years, putting tremendous
pressure on drug manufacturers to expend capital when the risk of drug failure is still high. Existing
manufacturing plants costs linger between $10 50 million depending upon required output and
therapeutic bioproduct. At the two extreme price ranges, a 100 L, $10,000 monoclonal antibody
plant scales up to a 10,000 L plant costing $120 million. The value of the primary and secondary
US biomanufacturing market in 2006 is estimated to be approximately $50 billion with a healthy
growth rate almost approaching double digits due to the escalating number of biotech drugs in
the pipeline and skyrocketing sales of approved high value, life saving biopharmaceticals.

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Biodisposables

Chapter 5: BiodisposablesFacility Design

Developments

and

Future

Facility Design Capabilities with Disposables

Single-use technologies are not only available for an increasing range of applications, but
they are also expanding from stand-alone devices to multi-component systems. With this
growing trend comes a greater number of benefits and implementation considerations.
Contract manufacturing organizations (CMOs) and biotech start-ups were among the earliest
adopters of disposables technology. CMOs saw them as a way of minimizing the risks of crosscontamination, whereas new biopharmaceutical companies enjoyed a considerable reduction
in the need for capital investment.
Both groups recognized the significant time and cost benefits of eliminating cleaning, cleaning
validation, and steam sterilization validation, as well as the reduced potential for operator
error. Today, the single-use model is attracting new converts, particularly in the area of patientspecific therapies. Given the small batches in which such therapies are processed, it is now
practical to move toward a totally disposable manufacturing paradigm. Availability of disposable
technologies for increasingly larger scale processes both upstream and downstream is reshaping
the way biopharmaceuticals are made and providing an efficient path to manufacturability for
emerging therapies.
Single-use technologies have gained increasing acceptance by the industry as a means of
achieving safe, compliant, and efficient processes. As the industry adopts to the technology
benefits, there is a demand for further integration of disposables into processes. The goal is
to develop integrated single-use solutions that offer certain process functionalities (such as
mixing, temperature control, aseptic connection, disconnection, and controlled freezethaw)
within a closed system. Recently there has been a strong emphasis on changing the mindset in
the biotech industry by moving the focus to process optimization and operational efficiency as
a way to reduce overhead.
An expansive and growing array of disposable technologies exists for downstream processing
applications, including direct-flow filtration (DFF) filters, tangential-flow filtration (TFF)
cassettes, membrane chromatography capsules, lenticular depth filters, filling equipment,
aseptic connections devices, tubing, adaptors, clamps, and bags. The availability of this array
of disposables technology has catalyzed new thinking with regard to processing approach and
facility design.
As a closed-loop system, it prevents the need to disassemble, transport, clean, validate, and
reassemble components in a classified cleanroom environment. In many cases, disposable
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Biodisposables

products are supplied pre-sterilized to eliminate the need for steam-in-place (SIP) or autoclaving.
Opening a package and plugging a single-use device or multi-component disposable system
into a product train offers a simplification to process development staff that otherwise must
develop extensive cleaning protocols. The result is not only labor savings, but also a shift in
facility design toward fewer cleanrooms and reduced environmental monitoring requirements.
With single-use operations, applications no longer need to be physically segregated. Instead,
they can be performed side by side as closed-loop systems. This allows for the more efficient
use of facility space, especially for CMOs and biotech start-ups.
Even as disposable systems seal off processes from contamination, their translucency provides
operators with visibility into manufacturing operations. Users can observe flow and fluid levels
as well as spot fluid discoloration and air pockets immediately. Single-use processes also allow
for a high degree of modularity in that capacity can be built out gradually in phases as demand
increases. Within conventional facilities, not only do hard-piped systems need to be factored
into the initial facility design, but also oversized utility systems are specified in anticipation
of future needs. Retrofitting is significantly easier with disposables than with fixed equipment
since disposable components are inherently modular.
Disposables can also simplify transfer of the drug production processes to other manufacturing
sites, such as CMOs, or other facilities within a company. Also, without the need for significant
capital investment in hard-piped systems, single-use technologies have allowed biotech startups to manufacture their products in-house. This option provides these companies with more
control over the development process and enables production to be accelerated as needed.
Previously, outsourcing production was the only economical option. This new flexibility enables
companies to better manage their own manufacturing expenses and investments during later
development stages, when greater drug supply requirements for advanced clinical trials can still
carry considerable risk of product failure.
The FDA has been a major proponent of disposables technology. Considering that validation
accounts for 1020% of the cost of a new plant, disposables provide the industry with practical
solutions to alleviate the challenges and costs associated with cleaning operations. Singleuse (disposable) manufacturing technology can be particularly useful in clinical phases of
development, when a company may handle a variety of drugs each year with limited quantities
required. The burden of conducting necessary changeovers and associated validation studies
when using stainless steel equipment reduces available production time in a multi-product
facility. Disposables may be substituted in numerous unit operations in a biological production
process such as production (bioreactors), mixing, product transfer, connection/disconnection,
filtration, chromatography, centrifugation, storage, sampling, and filling.
Exhibit 5.1 provides design consideration steps when considering the use of disposables in a
single or in multiple biomanufacturing unit operations.
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