Cambridge Healthtech Institute’s 2nd Annual
Advances in Purification Technologies
Novel Advances and Continuing Improvements for Higher Downstream Productivity and Purity
August 6-7, 2015
Part of CHI's 7th Annual The Bioprocessing Summit
August 3-7, 2015 | Westin Copley Place Hotel | Boston, Massachusetts
The downstream processing field is experiencing a resurgence of vitality and innovation. Driven partly by the quest for higher titer and higher throughput for existing mAb processes and partly by the next generation, non-monoclonal molecules coming down
the pipeline, we are seeing exciting new developments in each step of downstream processing - from capture to purification to polishing. New, higher capacity and multi-modal ligands that can eliminate aggregates; advances in filters and flocculants
for better binding and getting rid of impurities; process improvements from 3 columns to 2 columns and the promise of continuous purification technologies - all these are testaments to the exciting innovations that aim to deliver higher purity, higher
titer and higher throughput.
CHI’s Second Annual Advances in Purification Technologies will invite scientists and engineers to share their experiences, trials and results with these new technologies and continuing improvements, where they have encountered
challenges, and what pitfalls to avoid.
Thursday, August 6
1:55 pm Chairperson’s Remarks
Yi Li, Ph.D., Sr Scientist, Biologics Process Development, Bristol-Myers Squibb
2:00 KEYNOTE PRESENTATION
How to Scale-Up Precipitation of Recombinant Antibodies
Alois Jungbauer, Professor, Department of Biotechnology, BOKU and Head of Bioprocess Engineering,
Austrian Centre of Industrial Biotechnology
Precipitation and flocculation has been considered as potential methods for capture of antibodies especially when perfusion culture is used for production. Ethanol precipitation and precipitation by non-charged polymers reached equilibrium in less
than 1 minute. As rectors for such fast operations tubular reactors have been studied. As scale-up criteria fractal dimension of the flocks have been used. Mechanical compression during harvesting is influenced by dissolution kinetics.
2:45 Fatty Acid Reduction of Chromatin Heteroaggregates from Mammalian Cell Culture Harvests
Wei Zhang, Ph.D., Research Scientist, Downstream Processing, Bioprocessing Technology Institute, Singapore
Saturated and unsaturated fatty acids ranging in chain length from 6 to 12 carbon atoms were evaluated over a range of pH and conductivity values to identify the combinations supporting the most effective reduction of aggregates, product-related impurities,
and host contaminants while conserving high recovery of native IgG. Sensitive analytical methods were also developed. Results will be presented tracking contaminants and fatty acids through several 2-step IgG purification processes.
3:15 Development of Robust Impurity Clearance Methods Using Chromatographic and Non-Chromatographic Techniques
Srinivas
Chollangi, Ph.D., Scientist II, Biologics Process Development, Bristol-Myers Squibb
3:45
Production Scale Pre-Packed Chromatography for Use with 1000L & 2000L Bioreactors
Christine Gebski, Vice President,
Product Management, Repligen
How do we scale pre-packed chromatography to production-scale? The OPUS® 45 and 60 cm columns can purify a feed stream from 1000L - 2000L bioreactors. We will show how the unique design of these GMP-scale columns deliver equivalent or better performance
as self-packed glass columns, and demonstrate how usage can result in substantial time and cost savings.
4:00 Ice Cream Social in the Exhibit Hall with Poster Viewing
4:45 High Pressure Refolding as an Alternative Technology for the Manufacture of Biopharmaceuticals
Linda Gombos, Ph.D., Postdoc, Institute of Biotechnology (BOKU), Process Science Downstream Processing, BOKU-Boehringer Ingelheim RCV
We evaluated the application of high hydrostatic pressure for refolding a variety of biopharmaceuticals. Our aim was to systematically compare this emerging technology to conventional refolding and to identify its potential economic benefits. We also
compared the structure and activity of the two protein variants to address concerns on therapeutic effect and safety. Our results indicate that high-pressure is a potent technology for refolding proteins with significantly higher productivity and
in unaltered quality compared with conventional refolding techniques.
5:15 Investigation of Fouling Mechanism of Hydrophobic Interaction Chromatography Resin during Monoclonal Antibody Purification
Theodore
T. Diakov, MSc, Associate Scientist, Downstream Process Development, Bristol-Myers Squibb
Fouling of chromatography resins during repeated cycles of use can be a significant problem as it can compromise the capacity to clear impurities as well as cause inconsistent performance. Identifying the fouling species and mechanism can provide valuable understanding and assist in designing more efficient cleaning strategies. In this work we examined fouling of hydrophobic interaction chromatography resin during a monoclonal antibody purification process. Historically, HIC resins have been shown to allow protein unfolding on the surface under favorable conditions and we propose that protein deposits adsorbed by this mechanism are exhibiting resistance to traditional cleaning approaches. We report our comparisons of the resin performance in fouled and non-fouled state, the results of our pursue in identifying the nature of the foulant and its localization within the resin and our approaches in cleaning.5:45 End of Day
6:30 - 9:00 Recommended Dinner Short Course
Friday, August 7
8:00 am Registration and Morning Coffee
8:25 Chairperson’s Remarks
Gregory Zarbis-Papastoitsis, Ph.D., Vice President of Process and Manufacturing Sciences, Eleven Biotherapeutics
8:30 Modular Approaches to Express and Purify Complex Therapeutic Proteins
Stefan Schmidt, Ph.D., Vice President DSP Production, Rentschler Biotechnologie GmbH
Difficult to express proteins represent an increasing amount of therapeutic molecules. This causes bioprocessing challenges such as controlling glycosylation, increasing titers and yields and suppressing aggregation. Here I demonstrate how to develop
modular processes based on DoE principles solving these issues. Various case studies highlight successful process design, optimization strategies and critical manufacturing parameters. Additionally practical advice will be given on what to consider
when designing a novel molecule.
9:00 Single Step Purification of Biotherapeutic Proteins in a Highly Simplified Process Using an Optimized, Proprietary Fragment Complementation System
David O’Connell, Ph.D., Lecturer, Director, MSc Programmes, Biotechnology, School of Biomolecular
& Biomedical Research, University College Dublin
Engineering of a very high affinity complementation between two calcium-binding protein domains has led to the development of a robust purification system that in a single step produces high yields of very pure protein for downstream analysis, dispensing
with time consuming dialysis step. Through collaborations with leading biopharmaceutical companies and research institutes the purification of proteases, kinases and binder proteins will be presented.
9:30 Assembly of Knob and Hole Bispecific Antibodies
Josefine Persson, Ph.D., Senior Group Leader, Purification, Genentech, Inc.
We have produced glycosylated and aglycosylated CHO and E. coli Knob & Hole Bispecific Antibodies. In all cases the assembly conditions and assembly effectiveness are very similar. Our data shows that the assembly step is very robust and efficient
in producing the Knob & Hole Bispecific Antibodies while minimizing the amounts of homodimers (mAb).
10:00 Coffee Break
10:45 The Challenges of Developing Processes for New Protein Formats
Andreas Schaubmar, Ph.D., Head of Downstream Processing, Large Molecule Research, Roche
Pharmaceutical Research and Early Development, pRED, Roche Innovation Center Penzberg
Innovative biologics often combine several protein domains in increasingly complex formats to achieve superior therapeutic effects. We developed manufacturing processes for several new formats that provide the required yield and purity for clinical studies.
For that we considered the design of the therapeutic format and integrated the selection of a suitable cell line, the development of a productive fermentation process and a complementary purification scheme in a process effectively minimizing product
related impurities.
11:15 Scaling-Up of a Downstream Purification Process for a New Recombinant Product (rhFVIII)
Martin Linhult, Ph.D., Head of Bio100 line 1, Octapharma
Octapharma has developed a new process for the production of a recombinant human FVIII product derived from a human cell line (HEK293F cells). Clinical trials are ongoing with positive results. During process development several different approaches have
been tested, old established techniques as well as new ones has been evaluated. In this presentation I will discuss scale-up of a new downstream purification process and also different affinity ligands that could be applied.
11:45 Purification of a Highly Disulfide Linked Protein by Cation Exchange Chromatography
Thomas Linke, Ph.D., Senior Scientist, Purification Process Sciences, MedImmune
We present an informative case study on the development of a direct capture step of a refolded, novel biotherapeutic by ion exchange chromatography without prior buffer exchange. High yield and product purity in a single step made it possible to replace
two affinity columns used in the pre-clinical manufacturing process. Low solubility in solution and a propensity toward aggregation presented special challenges for process development.
12:15 pm Sponsored Presentation (Opportunity Available)
12:30 Luncheon Presentation: Preventing Virus Contamination of Bioreactors with a Virus Barrier Filter
Christina Carbrello, Ph.D., Senior Scientist, Process Solutions, EMD Millipore
Cell culture media is typically sterile filtered to remove bacteria and mycoplasma, however many bioreactors remain unprotected from viral contamination. Ideally, a filter could be used to remove viruses, but filtration is traditionally perceived
as being unsuitable for the upstream processes. A new virus filter has been evaluated and demonstrated high levels of virus, bacteria and mycoplasma removal while providing high flow and capacity in chemically defined media with little to no impact
on cell culture performance.
1:15 Session Break
1:25 Chairperson’s Remarks
Julie Bomholt, Ph.D., Protein Specialist, Research & Development, Aquaporin A/S
1:30 Membrane Proteins: From Lab-Scale towards Large-Scale Production
Julie
Bomholt, Ph.D., Protein Specialist, Research & Development, Aquaporin A/S
Downstream processing of membrane proteins is further complicated by the amphipathic nature of membrane proteins that even in the purified form are ternary complexes consisting of protein, lipid and detergent in unknown ratios. By use of the physiologically
important trans-membrane protein family of aquaporins as model proteins, we developed a S. cerevisiae-based expression system and identified conditions that allowed us to substantially increase the membrane density of recombinant functional
aquaporin.
2:00 Antigen Purification for Biologics Projects
Medha
J. Tomlinson, Ph.D., Principal Research Scientist, Global Biologics – Protein Sciences, Abbvie Bioresearch Center
The early discovery stage of antibody programs requires generation of proteins and cell lines. Appropriate construct design and expression systems are needed to retain antigen biophysical characteristics and activity. Proprietary antibody generation
requires expression and purification of various orthologues for screening and optimizing affinity. Rapid turnaround and antigen supply are paramount for success. This talk will focus on working towards building a robust platform to overcome
many of these challenges.
2:30 Miniaturized and Automated Inclusion Body Processing – From Homogenization to Purification
Cornelia
Walther, Ph.D., Post-Doc, Institute of Applied Microbiology/Biopharma Austria Process Science, University of Natural Resources and Life Sciences Vienna
Biopharmaceuticals overexpressed in E. coli are often deposited as high density aggregates in inclusion bodies requiring additional process steps such as inclusion body recovery, solubilization and refolding. For each product, individual needs
of the process might present a bottleneck during downstream processing. We present a miniaturized platform for parallel screening of cell disruption, IB washing, solubilization and refolding conditions and purification compatible with the
concept of DOE.
3:00 Networking Refreshment Break
3:15 Scaled-Down Model Qualification and Use for Process Improvement
Adam J. Meizinger, Process Engineer, Purification, Manufacturing Science and Technology Laboratory,
Genzyme, A Sanofi Company
3:45 Challenges in Developing Predictive Downstream Scaled-Down Models
Omkar Joshi, Ph.D., Head, Downstream Development, Global Biologics, Bayer
Healthcare
Scale-down models as appropriate representation of the manufacturing process are indispensable tools in characterization of biopharmaceutical manufacturing processes. During process development, such models enable evaluation of variability in input materials and parameters on a process and its impact on product quality to an extent that simply is not feasible at manufacturing scale. The key is to keep it simple and appropriate to needs. The presentation shows a pragmatic and systematic approach how to increase understanding of an antibody purification process by elements like scale-down modelling and qualification, cross-functional risk assessments, and process characterization studies.
4:15 Close of Conference