Preview: European Biotechnology News Special 6-7/2013 - Single Use Equipment

European Biotechnology News Science & Industry

June 2013

II Single-Use Equipment

SPECIAL 29_EBN6_13_SPECIAL_titel_tg.indd 29

07.06.2013 10:32:25 Uhr

European Biotechnology Net work

Join the European Biotechnology Network!

The European Biotechnology Network is dedicated to facilitating co-operation between professionals in biotechnology and the life sciences all over Europe. This non-profit organisation brings research groups, universities, SMEs, large companies and indeed all actors in biotechnology together to build and deliver partnerships.

Do you want to know more about the advantages of a (free) membership? Just have a look at our website: www.european-biotechnology.net

European Biotechnology Network | Avenue de Tervueren 13 | 1040 Brussels, Belgium Tel: +32 2 733 72 37 | Fax +32 2 64 92 989 info@european-biotechnology.net | www.european-biotechnology.net

30_EBSIN6-7_13_EBN.indd 1

05.06.2013 17:33:45 Uhr

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

31

Single-Use Equipment Intro

Disposables going stronger by the day Unprecedented growth in biologicals and cell therapies is also powering the market for the single-use equipment employed in cell culture, drug discovery and biomanufacturing.

According to a recent analysis released by the Boston Consulting Group, sales of biological drugs grew 8% in 2012, while the research market for single-use cell culture vessels ballooned 17%. Bioplan Associates predicts the GMP biomanufacturing market for disposables like plastic bags or fermenters will quadruple to US$2.8bn by 2016.

2000L, filters and other components. “Experts expect that fermentation volumes of maximum 2,000 liters will be the norm in future generations of bio­pharmaceuticals,” says Uwe Gottschalk. The Vice President of Purification Technologies at Sartorius Stedim Biotech says that’s down to the increased effectiveness of batches of mammalian cell culture produced in this volume range, as well as smaller patient populations resulting from individualised treatment standards and improved manufacturing methods. “Such manufacturing systems can be more quickly engineered as single-use unit operations,” believes Gottschalk, “and – because of their high proportion of variable components – cost money only when they are actually in

© University of Wisconsin

“We’ve seen an extension in cell culture as the industry looks more towards the development of biologics from cultured cells,” says Richard Eglen, VP and General Manager at Corning Life Sciences, the market leader in the area of small-scale disposable cell culture vessels (see p. 36). “This area of bioprocessing is very close to cell culture.”

The advent of cell therapies and stem-cellbased drug discovery is expected to generate further growth in the research market for single-use culture vessels, microplates, tubes, and roller bottles in the future.

The same is true on industrial scales, particularly when it comes to disposable biomanufacturing equipment such as storage bags, fermenters with volumes of up to

A coated single-use culture vessel used in stem cell cultivation

31_EBSIN6-7_13_Special_intro_tg.indd 31

operation.” Until recently, most single­-use bioreactors were only suitable for mammalian cell culture. However, a brand-new case study performed by Eppendorf/DASGIP shows that they can also be used in bacterial fermentation (see p. 38).

With a current market share of just 10%, single-use bio­manufacturing systems are still predominantly used in bio­pharma R&D. Bioplan Associates says that is about to change. More and more pharma companies are outsourcing biomanufacturing, and the Contract Manufacturing Organisations (CMOs) that use single-use equiment are seeing an annual growth rate of almost 15%.

That’s largely due to reduced capital investment compared to stainless steel fermenters, improved flexibility, the decreased risk of cross-contamination, and quicker turnaround times as clean-in-place requirements become redundant. However, the success story in upstream processing for single-use suppliers such as Thermo Fisher Hyclone, GE Healthcare/Xcellerex, Sartorius Stedim or Merck Millipore has not been transferred to downstream processing of bio­logicals.

Growing need for DSP solutions Vastly improved cell culture product yields are threatening to swamp biomanufacturers’ downstream capacities. “The need for advanced biomanufacturing is growing critical,” says Sebastien Ribault, Director Bioproduction & Development, Bio­Pharm Process Solution at Merck Millipore (see p. 40). He and his colleages evaluated over 10 capture technologies and more than 40 flow-through purification chemistries to create an optimal downstream processing template for continous processing of mono­clonal antibodies. In this issue, Swiss Filtrox also presents the very first depth filter to use the advantages of the alluvium technology in a disposable format.

Despite advances in the single-cell area, challenges remain. A ring test evaluating the influence of ten different bag materials on six mammalian cell lines showed two of them negatively influenced cell growth.

Standards to prevent such effects of solvents leaching from the plastic are urgently needed, say the researchers from DECHEMA’s single-use working group.

B

07.06.2013 10:32:44 Uhr

32

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

single-Use Equipment Interview

Biopharma needs more innovation Increasing cost pressures are prompting the pharma sector to explore new approaches to manufacturing. EuroBiotechNews spoke with Uwe Gottschalk, Vice President of Purification Technologies at Sartorius Stedim Biotech, about current trends in bioprocessing technology.

Euro|BioTech|News

?

Sartorius Stedim Biotech has grown continuously for years, and presented strong figures for both 2012 and the first quarter of 2013. What drove growth?

!

Gottschalk: The major growth drivers have been our single-use products, which are increasingly being used as alternatives to reusable stainless steel equipment in the production of bio­pharmaceuticals. Early on, we identified this sustainable trend toward single-use technologies, and moved quickly to build up a broad-based portfolio of products. We are now benefiting from those. Over the medium and long term, our business will also continue to be fueled by stable trends. The global population is growing and living longer, and millions of people in emerging economies are for the first time gaining access to healthcare. Moreover, the trend is shifting from conventional chemically synthesised medications to biologics, and this will continue in the future.

products at ever-lower prices, while retaining the same expected high quality. In effect, this means our customers are seeing a margin squeeze as a result of these two aspects. A visible sign of this development is the arrival of biosimilars and biobetters.

Based on available regulatory drafts, these are expected to receive approval – in the US as well. This is why many established pharmaceutical companies have entered into alliances in this area. Amgen surprisingly announced at the beginning of this year that it was getting involved in the development of generic versions of top-selling bio­tech drugs, and would therefore be competing head-to-head with large innovator companies such as Roche, Abbott and Merck. For me, that was an amazing announcement. It was like when the former Pope said he was resign­ing. In the final analysis, the competition is heating up, markets are becoming fragmented and the pressure on manufacturing processes is growing.

Euro|BioTech|News

?

What trends do you forecast in markets for upstream and downstream processing?

Single-use systems are increasingly replacing conventional stainless steel bio­reactors.

Why is the traditionally conservative pharma sector relying on these innovations?

!

Gottschalk: Our customers’ competitive environment in biopharmaceutical manufacture is currently undergoing sweeping changes. In addition to rising costs and the known risks entailed in developing new drugs, pressure is mounting on our customers to manufacture such

!

Gottschalk: Given the number of bioreactors that have been installed, the fermentation capacity available worldwide for mammalian-cellbased production is currently estimated at about three million liters. Considering the productivity increases attained over the

Euro|BioTech|News

32-34_EBSIN6-7_13_Special_Gottschalk_tg.indd 32

?

A recognised expert for downstream process technology, Dr. Uwe Gottschalk holds a degree in chemistry and is the Vice President of Purification Technologies at Sartorius Stedim Biotech. Before joining the firm, he worked from 1991-2004 in various positions at Bayer Healthcare, where he was in charge of purification of monoclonal antibodies and recombinant proteins in various expression systems.

past years, we can already speak of this as overcapacity. At the same time, we are experiencing a boom in single-use systems.

This contradiction can be resolved if you compare the particular cost base structures. Stainless steel systems require high capital expenditure, and their operation is dominated by fixed costs. For this reason, they are the better choice for longterm market supply for large quantities of pharmaceuticals, and the top 10 antibody products fall within this category. For the future generation of bio­pharmaceuticals, experts consider fermentation volumes of 2,000 liters maximum will be the norm. The reasons they cite, for example, are the increased effectiveness of batches produced in this volume range, or smaller patient populations resulting from individualised treatment standards and improved manufacturing methods. Such manufacturing systems can be more quickly engineered as single-use unit operations and – because of their high proportion of variable components – cost money only when they are actually in operation. That’s an attractive option, especially for small companies.

At the same time, the innovative technologies implemented in single-use systems

05.06.2013 17:34:32 Uhr

26367 EBN Ad. 30/05/2013 11:38 Page 1

Experience Flexibility Cell culture - Gene to GMP Following recent significant expansion of our mammalian cell culture facilities we offer a flexible approach to program delivery, for your mAb, ADC or non-mAb: l Cell line development, using in-house, CHO-based

expression system l HTS enabled process development, transfer and

characterization l cGMP cell banking

l cGMP manufacture in new, purpose built flexible facilities

up to 2000L, including the latest single-use technologies l Full analytical development for novel biologics and

biosimilars l Choice of US or UK production sites.

www.fujifilmdiosynth.com

33_EBSIN6-7_13_Fujifilm.indd 1

06.06.2013 12:40:52 Uhr

34

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

Single-Use Equipment provide product-differentiation benefits and competitive advantages

Euro|BioTech|News

?

In what directions can increased productivity in upstream processing be further optimised?

!

Gottschalk: Because of the substantial increase in the performance of mammalian cells, largevolume production systems will be less frequently used in the future. While it is hardly feasible to further increase the expression rate per cell, there is still considerable potential for optimising biomass density, for example, in comparison with microbial systems. Looking at values upwards of 100 g/L for secondary metabolites, we can probably expect anti­body titers to con­tinue to rise.

However, in our drive to further optimise upstream processing, we still need to continue giving priority to quality factors such as processability over pure biomass growth.

Euro|BioTech|News

?

The burst of productivity poses major challenges for downstream processing. What developments can we expect there?

!

Gottschalk: Unfortunately, what I have said so far does not apply equally to downstream processing. Here we think in terms of mass, not volume. While you can produce a much higher protein yield using a given bioreactor volume, downstream processing of this yield requires correspondingly larger chromatography columns, including their peri­ pheral components. Although we are seeing a certain productivity gain in downstream processing as well, it is not enough to fill the gap that’s been created. Besides action taken to ensure continuous improvement and operating excellence, only truly innovative technologies will help lead to quantum leaps in productivity. Examples that come to my mind in this context are the further consolidation of process steps and the introduction of integrated, continuous processes.

Euro|BioTech|News

?

What role will improvements in process development and validation play?

32-34_EBSIN6-7_13_Special_Gottschalk_tg.indd 34

!

Gottschalk: In the early phase of pharmaceutical biotechnology, production facilities resembled “venerable” factories in which efficiency did not play a role. R&D processes were simply scaled up, and that was it. Real industrialisation of processes, along with enhanced productivity levels, simply did not take place.

Manufacturing procedures were hands-off when it came to rethinking workflows, and biotechnology seemed to play a distinct role in comparison with other sectors of industry. Changes were frowned upon, and could only be reluctantly pushed through in – at most – small increments. Since then, the barriers to the introduction of new technologies have also been lowered for quite some time as a result of initiatives taken by regulatory authorities – among them the Quality by Design (QbD) approach, and the endorsement of process analytical technology (PAT). Manufacturers without access to new technologies will increasingly come under regulatory and economic pressure.

Even the production of biosimilars will not require any replication of the original manufacturing process as long as the specifications of the final products are met. Overall, the tide is turning. The ideal approach is not to cling to time-worn principles that presumably minimise risk, but to introduce the latest competitive standards as soon as these prove to be robust enough for GMPcompliant operations.

Euro|BioTech|News

?

of sterile biopharmaceuticals. Such standards will consequently become widely accepted solely by virtue of drug safety. Apart from this, single-use devices happen to reduce costs and, on closer analysis, are the better choices for managing and conserving natural resources. I personally expect that in the future, biopharmaceutical manufacturers will have to justify why they reuse products in critical process steps like viral clearance. The same also goes for the respective chromatography steps.

Euro|BioTech|News

?

What will the more open attitude of biopharmaceutical manufacturers toward innovation mean for your business?

!

Gottschalk: In some areas, we fought to gain acceptance of innovative methods, such as the use of membrane adsorbers for the removal of product contaminants. Now these are good examples of disruptive innovations in the conservative market of biopharmaceutical production. Single-use bioreactors are further examples of a trendsetting innovation.

Still, it would be presumptuous to state that the trend toward single-use products was our doing. Rather, this current development is driven by a market need, and technology providers are hard-pressed to keep pace with it. But we embrace such challenges, which we can definitely solve.

Euro|BioTech|News

?

Currently we are still far from talking about a fully single-use process chain. Are singleuse technologies still niche solutions?

What about your growth strategy? Is it based on organic growth? Or are you also planning acquisitions?

!

Gottschalk: The biopharmaceutical industry is certainly not among the trailblazers when it comes to using innovative technology. But there is tremendous potential out there for this industry to upgrade equipment employed in critical process steps to single use. Take the medical sector as a comparative example. When was the last time you received an injection with a reusable syringe? And would you even be willing nowadays to let your doctor come near you with such an old-fashioned device?

This illustrates that similar, modernised standards should apply to the manufacture

!

Gottschalk: For 2020, we are projecting that Sartorius Stedim Biotech will earn sales revenue of D1.5bn. In other words, we are striving to nearly triple our business. We are now in the fortunate position of having many organic growth opportunities available, primarily in North America, where we are predominantly striving to expand our market share. Then there’s China, where we will participate in building up the biopharma industry. However, further acquisitions and alliances are also conceivable, provided they are a good fit with our technology portfolio. 

07.06.2013 10:38:25 Uhr

Designed for DASbox® parallel mini bioreactor system

Single-Use Simplicity Our new BioBLU® 0.3f vessel for fermentation The BioBLU 0.3f for microbial applications perfectly combines the advantages of single-use solutions with a scalable stirred tank design and parallel process performance. With this fully instrumented mini bioreactor all critical process parameters can be precisely controlled.

> Small working volumes of 65 – 250mL > Magnetically coupled overhead drive safely operates up to 2,000rpm > Easy to use and fully instrumented for advanced process development > DoE available with DASware® software

www.eppendorf.com DASGIP ®, DASware ® and DASbox ® are registered trademarks of DASGIP Information and Process Technology GmbH, Juelich, Germany. Eppendorf ®, the Eppendorf logo ® and BioBLU® are registered trademarks of Eppendorf AG, Hamburg, Germany. New Brunswick™ is a trademark of Eppendorf AG, Hamburg, Germany. All rights reserved, including graphics and images. Copyright ©2013 by Eppendorf AG.

ebn_epp_ad_210x275_2013_06_07.indd 1 35_EBSIN6-7_13_Eppendorf.indd 1

06.06.201308.05.13 12:41:1012:50 Uhr

36

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

Single-Use Equipment Corning Life Sciences

Systematic expansion Driven by tremendous growth in the drug discovery of biologicals and cell culture research, Corning Life Sciences is making good on its stated goal of becoming a billion dollar business by 2014. Last October, the US-based provider of single­-use plasticware, media, and surface chemistry – as well as drug discovery products and services – completed the acquisition of one of the biggest players in the cell culture market, paying US$720m in cash for the majority of Becton Dickinson’s (BD) Discovery Labware business.

Even before the full integration of the co-leading supplier of single-use plastic consumable labware, the acquisition will expand Corning Life Sciences revenues by roughly 40%. BD Discovery Labware’s business unit sales hit US$289m last year, while Corning took in US$657m – almost twice as much as in 2009 (US$366m). “As a glassware provider in academic and drug discovery research circles, Corning’s history in the

life sciences stretches back around 90 years, but our business has particularly developed in the last five,” says Richard Eglen, Vice President and General Manager at Corning Life Sciences. “Since 2009, we have generated added growth through highly selective acquisitions that expanded our drug-discovery and cell-culture business, as well as our geographic footprint.” According to Eglen, products from those two areas contribute

BD Discovery Labware product lines integrated into Corning Life Sciences under the same brand name Brand name

Product

A Corning BioCoat® - (Corning Matrigel, Poly L-lysine, Poly D-lysine, Collagen, Fibronectin, Laminin)

Biological surfaces for improving cell attachment of adherent-growing cells

A Cell-Tak™

Cell and tissue adhesive

A Falcon®

Cell culture plasticware including tubes, cell culture flasks, inserts etc.

A FluoroBlok™

Fluorescence-blocking high density PET membrane inserts for multiwell plates

A Nu-SerumTM

Low-protein growth medium supplement

A Primaria™

Cell culture plasticware and microplates

A Corning PureCoat™

Amine and carboxyl surfaces for the culture of adherent cells

A Sero-LinerTM

Centrifuge disposable plastic insert

A Corning Gentest Supermix® / Corning Gentest™

Tissue fractions, P450/UGT enzymes, Corning Supersomes™, transporters, and contract ADMET research services

A Epranex®

Heparin binding plate designed to immobilise heparin without modification

A Gold Seal®

Microscope slides

A Supersomes™

Recombinant drug-metabolising enzymes & controls

A UltraPool™

Human liver microsomes for use in in vitro ADMET drug discovery screening and development

36-37_EBSIN6-7_13_Special_Eglen_tg.indd 36

Richard M. Eglen joined Corning Life Sciences as Vice President and General Manager in June 2011. The molecular pharmacist previously led PerkinElmer’s (PE) biodiscovery business. During his career in the biopharmaceutical sector, Eglen has held executive management positions at F. Hoffmann La-Roche, Syntex Research, LJL BioSystems Inc, and DiscoveRx.

around 50% each to the business unit’s revenues. With a series of acquisitions, Corning has systematically filled gaps in both segments.

Its expansion began in 2009 with the acquisition of Axygen Bioscience, a provider of plastic consumables for genomics research (Axygen® product line). That added sales forces in Europe, China and the US. Next up was Plaslab SAS, the holding company of French Plastique Gosselin SAS, an established European supplier of disposable plastics for microbial quality control and testing in the food and beverages industry. Then Corning extended its cell culture product portfolio with the acquisition of Mediatech, Inc. Products include a full range of synthetic and classic media for mammalian cell culture.

Growing with industry trends With the new acquisition of BD’s Discovery Labware business, Corning Life Sciences has continued to expand in Europe and important Asian markets. In the process, it has also created a market-leading business for cell culture vessels and coatings that allow cells to grow – either by blocking them from unspecific attach-

06.06.2013 12:44:30 Uhr

C M E

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

37

Single-Use Equipment ment through hydrophilic coatings (Corning’s Ultra-Low ® hydrogel) or by supporting cell anchorage to functionalised micro­­-carriers (Corning Biocoat ® ECM coatings). “We now have a larger range of surfaces for our different vessels, which links the different geo­metry to a range of different chemistries,” stressed Eglen.

Experts estimate that its combined market share in this area is just under 40%.

Besides BD Discovery Lab­w are’s plastics consumables and bio­logical surfaces (see table), which complement Corning Life Sciences’s offerings of chemically modified plasticware, three other product platforms will strengthen its drug discovery business: liquid handling products, cell-based assays and products for ADMET research.

One factor pushing Corning Life Sciences’ 17% growth over the last five years has been the R&D needs of the expanding bio­ logics industry: “We’re seeing an extension in cell culture as the industry looks

more towards the development of biologics from cultured cells,” says Eglen. “This area of bioprocessing is very close to cell culture. The other trend is that researchers tend to prefer technology providers that offer products covering the whole cellculture workflow of preparing cells, analysing cells, growing cells and using them in drug discovery.”

Set for the future Corning Life Sciences also recently widened its portfolio in another dynamic field: feeder-free culture of human embryonic and human induced pluripotent stem cells for drug screening. Until now, only biological coatings like Matrigel® allowed the attachment of the sensitive cells to microcarriers for stem cell expansion.

However, GMP-compliant manufacture will require chemically defined coatings that mimic the effect of extracellular matrix (ECM) proteins. Microcarriers coated

with the company’s new xeno-free synthetic surface Synthemax II supported expansion of functional undifferentiated stem cells in defined media for over 20 passages. The capability of automatically scaling up feeder-free expansion of stem cells using Hyperstack® cell culture vessels makes Corning Life Sciences a firstmover in the fast-growing area, providing the first commercially available system for culture of clinical-grade stem cells.

According to Eglen, directing stem cell differentiation in 2D-or 3D-cell cultures by specific coatings will be the next interesting field of innovation.

Geographically, Corning Life Sciences has already prepared for future growth. “We feel China is a very important market,” says Eglen. In late 2011, Corning Life Sciences opened a state-of-the art manufacturing and distribution facility in Wuijiang. Chinese market growth in the life sciences is expected to exceed 20% annually through at least 2015.

B

Contract Manufacturing Excellence Cobra invites you to visit our modern, purpose built, aseptic manufacturing facility in Matfors, Sweden.

Please visit our website to arrange a tour: www.cobrabio.com 36-37_EBSIN6-7_13_Special_Eglen_tg.indd 37

05.06.2013 17:42:18 Uhr

38

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

SINGLE-USE EQUIPMENT BACTERIAL FERMENTATION

Multifaceted solution Claudia M. Huether-Franken, Eppendorf AG, Bioprocess Center, Jülich

Single-use bioreactor solutions have been successfully established in both animal and human cell culture, and the technology is now making its way into microbial applications. In the following study, reproducible process control was achieved with single-use and reusable glass mini-bioreactors running in parallel. Fermentation of E. coli K12 led to reliable results, thus proving the tested rigid wall single-use stirred-tank vessels are an appropriate tool to accelerate microbial process development and shorten time-to-market.

Regardless of whether they are used for cell culture or in microbial applications, singleuse bioreactors provide several bioprocessing advantages in terms of time and cost effectiveness. Minimal setup times, no need for cleaning procedures and therefore reduced labor can accelerate bioprocess development tremendously. Compared to single-use bioreactors in cell culture, microbial applications make specific demands on bioreactor design and functionality. Fermentation processes need much higher kLa values for proper mass transfer, as well as suitable heating and cooling options. To

evaluate reliability in microbial fermentation processes when employing single-use technology, E. coli K12 (DSM 498) was cultivated in a fully-instrumented Eppendorf BioBLU® 0.3f single-use mini-bioreactor and compared to fermentations from conventional glass bioreactors.

Effective mass and heat transfer The rigid wall stirred-tank single-use bioreactor was specifically designed for microbial applications, and is equipped with a 2x Rushton impeller, liquid-free peltier

exhaust gas condensation and magnetic coupled overhead drive for high performance agitation. A four-fold parallel DASbox® Mini-Bioreactor System with active heating and cooling capacities was used with DASGIP Control Software for precise process control. Starting with a working volume of 0.1 L, the cultures were grown for 40 hours in PAN media with an initial glucose concentration of 40 g/L and fed with 50% glucose solution in the fed batch phase.

The temperature was controlled at 37 °C, and pH was adjusted to 6.8 via 4% ammonia solution. The cultures were submerged aerated through dip tubes with a constant rate of 6 sL/h (1 vvm). The dissolved oxygen was maintained at 30%, while stirrer speeds ranged from 600 rpm to 2000 rpm, which equal tip speeds of 0.94 m/s to 3.14 m/s.

The oxygen transfer rates (OTR) were automatically calculated via a DASGIP exhaust gas analysis module GA4. Highly reproducible OTR values of up to 250 mmol/(L·h) were observed in the single-use as well as glass bioreactors. Supporting kLa values of up to 2500 h–1 were determined by static sulfite depiction methods (data not shown) and demonstrate that this single-use bioreactor design perfectly matches the demands of microbial applications. The biomass production was determined offline as cell wet weight, and revealed comparable growth characteristics in single-use and glass bioreactors. The maximal biomasses of 160 g/L gained in the fermentations correspond to an OD600 of about 100 (data not shown).

Conclusion This case study shows that the BioBLU 0.3f single-use bioreactor addresses the specific needs of an E. coli fermentation, especially in regard to mass and heat transfer. Its specifically adapted single-use design supports the high demands of microbial applications.

Even though single-use bioreactor technology is currently mainly used in cell culture, it is also an effective tool to accelerate bioprocess development in microbial fermentation.  A. Reproducibility of online calculated oxygen transfer rates (OTR) of parallel process runs at constant temperature of 37 °C using single-use and reusable glass bioreactors respectively. B. Biomass production. Cell wet weight (cww) of fermentations carried out in single-use and reusable glass bioreactors.

38_EBSIN6-7_13_Special_DASGIP_tg.indd 38

Contact Claudia M. Huether-Franken huether.c@eppendorf.com

05.06.2013 17:41:22 Uhr

GE Healthcare Life Sciences Now part of GE Healthcare

Unlimited growth potential

Use one convenient bioreactor platform from process development to production.

Don’t let different vessel forms, materials, and control systems impede your growth.

Xcellerex XDR™ bioreactors simplify process scale-up and scale-down, providing a highly engineered, well-characterized product platform. Our service and support make scalability even easier. Spend your time growing cells — and outgrowing the competition.

www.gelifesciences.com/xcellerex

E: webinquiry@xcellerex.com P: 508.480.XCEL(9235) or 1.866.XCELLEREX

GE, imagination at work and GE monogram are trademarks of General Electric Company.

Xcellerex is a registered trademark of Xcellerex, Inc. XDR is a trademark of Xcellerex, Inc. © 2012 General Electric Company — All rights reserved.

First published September 2012.

1253076_EuroBiotechNews.indd 11 39_EBSIN6-7_13_Xcellerex.indd

10/5/12 9:44Uhr AM 07.06.2013 10:01:04

40

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

Single-use Equipment next Generation Downstream Template

rity. At these productivity levels, singleuse capture may be enabled through the use of very small columns that are cy­ cled many times to process one prod­ uct batch.

Continuous mAb processing

New adsorber technologies

Michael Phillips, Frederic Mann, Sebastien Ribault, Merck Millipore, BioDevelopment Center, Martillac, France

As the biopharmaceutical industry evolves, the need for advanced biomanufacturing is growing critical. Although many different manufacturing solutions have been investigated, most of the efforts have focused on three areas: (1) improving existing process templates through process compression to eliminate unit operations, (2) connected processing by combining unit operations, and (3) the introduction of technologies to enable fully continuous processing.

To meet the demands associated with these trends, Merck Millipore has devel­ oped a new processing template for mono­ clonal antibody (mAb) purification that: 1. Increases speed through the use of higher throughput unit operations re­ quiring lower cycle time 2. Decreases the cost of goods (COGs) 3. Increases manufacturing flexibili­ ty and facility utilisation by maximi­sing the use of single-use technologies 4. Minimises risk through the use of a ro­ bust template, enabling closed-process­ ing, and lowering upfront investments An integrated strategy was employed in developing the new template. Ultimate­ ly, over 10 capture technologies and more than 40 flow-through purification chemistries were evaluated. The optimal downstream processing (DSP) template comprised: – precipitation-based clarification with solids removal by depth filtration – continuous multi-column protein A chromatography capture – a fully-connected flow-through purifi­ cation train specifically developed for robustly purifying Protein A elution pools.

From a product capture perspective, Protein A chromatography was the only

40-41_EBSIN6-7_13_Special_Millipore_tg.indd 40

technology that met the objectives, ulti­ mately resulting in the lowest cost-ofownership. With the use of multi­-column continuous chromatography with incom­ pressible protein A resins, we were able to increase productivity more than 10fold without detracting from product pu­

Two new adsorber technologies were developed in order to support a robust flow-through purification train in postProtein A capture. The first new adsorb­ er was designed to remove host cell pro­ teins, leached protein A, and other small molecule additives to the cell culture me­ dium, including antifoam, surfactants, anti­biotics, and insulin. The second new adsorber was specifically designed to re­ move dimers and aggregates from mono­ mers in a flow-through mode.

These adsorber technologies were de­ signed to minimise the number of inter­ mediate solution adjustments required between steps, with only one pH change necessary. Consequently, several inter­ mediate pool tanks could be eliminat­ ed, resulting in a fully-connected flowthrough purification train. Additional­ ly, this train can be operated with only one processing skid, significantly re­

Conventional ConventionalProcess Process Conventional Conventional Process Process

® POD ® POD Centrifuge Millistak+ Bioreactor Bioreactor Centrifuge Millistak+ SHC SHC ® ® POD SHC Centrifuge Millistak+ Bioreactor Bioreactor Centrifuge Millistak+ POD SHC

Bioreactor Chrom Chrom Bioreactor Chrom Chrom

Conv Con

Centrifug Centrifu

Disposable DisposableProcess Process Disposable Disposable Process Process

Bioreactor Bioreactor Bioreactor Bioreactor

TMTM Clarisolve Clarisolve POD POD TM TM POD Clarisolve ClarisolvePOD

SHC SHC SHCSHC

Dis Di

Bioreactor Chrom Chrom Bioreactor Chrom Chrom

Clarisol Claris

Comparison of the conventional and the disposable process for mAB purification

07.06.2013 10:35:43 Uhr

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

41

Single-use Equipment ducing capital investment. A case study was conducted to exemplify the advantages of the new DSP process template through a direct comparison to a traditional template for the purification of a mAb from CHO cell culture. Experimental data indicate comparable yield, comparable host cell protein, DNA, leached protein A removal, and comparable product quality. However, the new process resulted in lower product aggregates in the final pool.

Processing more economically For commercial-scale operation, process modeling results indicate that the new process template would reduce the total manufacturing cost of goods by 11% at 1 g/L titer and 18% at 5 g/L titer. Most of these cost savings would result from significant reductions in capital investment, with the new process template consisting of more single-use technologies and fewer process skids and intermediate tankage. Moving to an entirely continuous process would result in an overall reduction of total manufacturing cost of goods by approximately 25%.

These additional reductions in COGs are driven by further reductions in capital, as well as lower labour costs.

A range of advantages In conclusion, Merck Millipore has developed a new process template for mAb purification that takes a holistic view of mAb processing. This template involves precipitation-based clarification, continuous protein A multi-column capture chromatography, and fully-connected flow-through polishing. Experimental data from a case study comparison has revealed that purification performance in the new process template is essentially equivalent to the traditional benchmark template. Process modeling results, however, also indicate that the new process template also brings significant economic benefits. Additional benefits for the new process template include lower processing time and a lower process footprint.

B

40-41_EBSIN6-7_13_Special_Millipore_tg.indd 41

ESBEs

New interest group for biochemical engineering takes first steps A Den Haag/Frankfurt – The manufacture and downstream processing of bio­logics yielded US$10.5bn last year, and according to Bio­p lan Associates, bio­p harma production and purification alone has been growing at a rate of 15%-18% annually for a decade. Wit­h the EU’s Horizon 2020 programme, another growth area is now gearing up for biochemical engineers who design and optimise processes involving biological systems: the biobased economy.

Growing with process development for the bioeconomy With tailwinds provided by huge EU funding budgets, biologists, computer scientists, engineers, and mathematicians have now joined forces to raise their profile, founding the European Society of Bio­chemical Engineering Sciences in Den Haag at the end of April. “The ESBES is nothing completely new,” says Guilherme Ferreira, the organisation’s first President-elect.

Founded in 1996 as a section of the European Federation of Biotechnology (EFB), membership figures are currently hovering around the 1,600 mark. “However, we felt that an independent organisation could give us better visibility at both the European and international scale.” The Portuguese bio­engineering and biosensor expert says the relaunched society covers several highly dynamic fields of innovation in the industry, among them new reactor designs, metabolic engineering and synthetic biology, downstream processing, process and pathway modeling and enzyme development.

“Natural” technology transfer Because half of ESBES members come from industry and the other half from academia, Ferreira believes ”building a common network for everyone involved

The first president of ESBES, Guilherme Ferreira, was elected at the founding meeting on 21 April 2013. in biochemical engineering will drive technology transfer on its own.”

Concrete plans for the society’s future activities will be made soon by its three funding societies: DECHEMA (Germany), IChemE (UK), and SFGP (France).

Some things have already been decided.

“We want to deliver new standards and bi­a nnual market reports,” says Ferreira.

High degree of expertise For policymakers, the new organisation pools a large number of top experts, which is urgently needed when drafting funding plans to make the bio-economy come to life. Currently the EU organisation pools its academic and industry know-how in five sections: downstream processing, bio­r eactor performance, biocatalysis, modelling – monitoring – measurement & control, and bioenergy systems.

Whilst the geographic focus of ESBES is currently central Europe, Ferreira sees further growth potential. “Our first task is to pool the expertise in bio­chemical engineering in the EU, which is dispersed throughout its different societies.”

B

07.06.2013 10:36:04 Uhr

42

Nº 6-7 | Volume 12 | 2013

Euro|Biotech|News

Single-use Equipment Microfiltration

Effective cell removal Ralph Daumke, Filtrox AG, St. Gallen, Switzerland

FILTRODISC BIO SD is a high performance single-use microfiltration system for removing cells and cell debris from fermentation broth.

module changes. Single-use methods are an ideal choice for contract manufacturers (CMO) and other operations that have a high frequency of product change. All systems can be delivered sterilised (gamma irradiation).

For alluvial filtration, the pharma grade DE (Celpure®, an Advanced Minerals brand) can be provided from FILTROX in singleuse transfer bags. The fermentation broth or cell homogenate is mixed with DE in a bio­reactor or mixing bag and then pumped over the filter unit. The DE and the cells/ cell debris form a filter cake, extending the system’s filtration capacity and preventing early blocking of the filter media. After filtration, the remaining liquid in the bag can be pumped over a FILTRODISC™ BIO SD 5” capsule to recover the last drops of the valuable liquid. After both filtrations have been carried out, the module and the capsule can be removed for disposal.

FILTRODISC™ BIO SD is also very useful for applications like active carbon re­moval or the filtration of highly toxic solutions.

Performance

Clarifications of fermentation broth and cell homogenate are one of the most impor tant steps in downstream biotech processes. Optimised fermentation processing leads to dramatically increased cell densities, accompanied by new challenges in clarification. Standard current technologies (centrifugation, depth filtration and membrane filtration) are no longer able to handle the high particle loads in an economical way.

Membranes are very cost intensive, and their scale up is impractical, as the required footprint for standard depth filtration increases with particle load. Centrifugation – an application in which mechanical stress is applied to cells – increases the turbidity through fine particles that have to be removed further downstream through separation and purification. Alternatively, a depth filter with higher capacity per area can solve these issues.

42_EBSIN6-7_13_Special_Filtrox_tg.indd 42

Allu­v ial filtration is a well-established method in pharmaceutical industries. Until recently, however, it was not used for cell separation as it was unavailable as a scalable and disposable system that fulfilled all validation requirements. FILTRODISC™ BIO SD is the first depth filter using the advantages of the alluvium technology in a disposable format, and is scalable from lab to industrial-size filtration.

Applications FILTRODISC™ BIO SD is a filtration system that removes particles (e.g. cells, bacteria, yeast) and other turbid matter (e.g. activated carbon) from process liquids like fermentation broths or cell homogenate.

The system is completely disposable. Single-use components reduce cross-contamination risks, eliminate cleaning validation efforts and reduce downtime during

The turbid matter in a cell broth is made up of cells, the feed and other degraded ingredients in homogenate from cell debris. Tests with E. coli bacteria (cell density 6.6 x 106), Pichia yeast (density 4 x 107) and cell homogenate were carried out to evaluate feasibility and obtain orientation values for this system. In every case, turbidity after filtration with 5” FILTRODISC™ BIO SD capsules was lower than it is after a standard centrifugation procedure.

Results from the 2” capsule trial filtration with E. coli could be directly used to filter a 600L batch without any other intermediate steps. For the production scale, a FILTRODISC BIO SD 16” double module was used.

Reusable stainless steel supporting shells allow users to fill the FILTRODISC™ BIO SD bags homogenously and protect them against pressure shocks. The heavy larger modules can be tilted horizontally to allow easy discharge of used bags. All DISCSTAR® BIO SD housings (except version 12K) are delivered with a separate trolley for module discharge. A pressure gauge is installed on the top of the support system.

B

05.06.2013 17:45:26 Uhr