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Overcoming Purification Challenges conference - Day 2

Archived Content

Overcoming Purification Challenges 

Day 1 | Day 2 | Download Brochure | Short Courses 

TUESDAY, AUGUST 21

8:00 am – 6:00 pm Registration and Morning Coffee

 

BREAKTHROUGH TECHNOLOGIES 

8:25 Chairperson’s Remarks

David W. Wood,Ph.D., Associate Professor, Chemical & Biomolecular Engineering, The Ohio State University

» 8:30 FEATURED PRESENTATION: 

Protein Adsorption and Transport in Polymer-Functionalized
Ion-Exchangers

Abraham-LenhoffAbraham Lenhoff, Ph.D., Allan P. Colburn Professor of Chemical Engineering, Chemical and Biomolecular Engineering, University of Delaware

Incorporation of hydrophilic polymers has become widely used by many adsorbent manufacturers in seeking increased capacity and improved performance of particulate, membrane and monolithic ion exchangers for use in bioprocessing. While such adsorbents are used in much the same fashion as their underivatized counterparts, the mechanisms involved and some features of the performance attained may differ in important ways.  This presentation will provide an overview of the structures of the derivatized materials and of similarities and differences in performance relative to their more conventional counterparts, with the goal of providing guidance for both adsorbent selection and design of new materials.
 

9:00 mAb-Aggregate Analysis in Two Minutes: Application of Tandem and Parallel Interlaced HPLC

Patrick-DiederichPatrick Diederich, Ph.D., Scientist, Institute of Process Engineering in Life Science, University of KarlsruheBiography 

Analytical techniques that match a high throughput process development approach are still not common or well characterized and often come along with lower precision. Quantitatively precise analytics such as HPLC techniques usually require long assay times. In this talk, methods are presented that show how HPLC analytics nevertheless can be utilized as high throughput compatible analytics. Most promising, the potential of combining interlaced sample injection with parallel, isocratic operation of two chromatography columns on a single HPLC system is demonstrated. By this methodology, the assay time for antibody aggregate analysis by size exclusion chromatography was reduced to less than two minutes per sample without sacrificing precision.

9:30 Rescue of Non-Expressing Protein Targets via Fusion to Maltose Binding Protein: Perspectives from a High-Throughput Protein Expression and Purification Pipeline

Steve-HewittStephen Nakazawa Hewitt, Ph.D., Senior Scientist, Department of Medicine, Division of Allergy and Infectious Diseases, School of Medicine, University of Washington - Biography 

Poorly expressing and insoluble protein expression are major obstacles for purification of recombinant proteins. We have developed several strategies for attempting rescue of these targets in a high-throughput manner in the context of a structural genomics effort. By modifying the cloning vector to include the highly soluble maltose binding protein expressed as a fusion tag, we are able to quickly attempt rescue of expression by simply moving PCR amplified inserts between various vectors.
 

Asahi Bioprocess10:00 Novel Hollow Fiber Membrane Adsorber, QyuSpeed™ D, Expands the Options Available in the Purification Tool Box

John Fisher, Senior Manager, Science & Technology, Asahi Kasei Bioprocess, Inc.

Membrane adsorbers are a powerful tool for polishing steps in downstream biopharmaceutical processes due to their high flux, ease of use, and simple scale-up. QyuSpeedTM D (QSD), a DEA ligand-grafted reusable hollow fiber adsorber provides high-throughput, robust impurity clearance over a range of pH values and salt concentrations. Scalability of this magnitude coupled with the ability of QSD to capture high molecular weight protein targets more effectively than existing anion exhange resins make QyuSpeedTM D the ideal choice for eliminating impurities from biotherapeutics or plasma-derived products.

10:15 Coffee Break with Exhibit and Poster Viewing

 

ANTIBODY PROCESSING 

11:00 Debottlenecking Antibody Manufacturing Through the Use of Simplified Buffer Systems

Nataj RamNatraj Ram, Ph.D., Senior Group Leader, Purification, Technical Operations, Abbott Bioresearch Center

One of the key materials used in the manufacture of proteins are buffers.  Buffer preparation requires a significant amount of space, time, and resources and could potentially be a bottleneck when dealing with high titer processes. In this presentation, the concept of using simplified buffer systems is introduced and demonstrated, as a means of reducing facility foot print, buffer volumes, buffer raw materials and possibly reducing manufacturing time.

11:30 Strategies to Streamline Two-Column Monoclonal Antibody Purification Platforms

Yun (Kenneth) Kang, Ph.D., Principal Scientist & Head, Purification Team, Bioprocess Sciences, ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company

In two-column mAb purification platforms, traditional Q column or, increasingly, Q membrane adsorber is used as a polishing step in a product flow-through mode. For mAbs with a low pI (< 7.0) or solubility issues under low ionic strength solution conditions, however, poor process performance is expected. We have developed a robust mAb purification platform which demonstrates high process yield and efficient clearance of impurities (HCP, HMW, host DNA, leached protein A) for those challenging antibodies.

12:00 pm Countercurrent Tangential Chromatography: New Single Use Technology for mAb Capture

Andrew ZydneyAndrew L. Zydney, Ph.D., Department Head and Walter L. Robb Family Endowed Chair, Chemical Engineering, The Pennsylvania State University - Biography 

Boris Napadensky, Vice President, Engineering, Chromatan Corp.

Countercurrent Tangential Chromatography (CTC) is a new column-free purification and capture technology that holds great promise for purification of high-value recombinant proteins like monoclonal antibodies. CTC provided high-resolution antibody purification at very low pressures (< 7 psi) with excellent protein recovery. In addition, CTC is no longer limited by the constraints of packed columns, allowing the use of smaller chromatography beads with improved binding kinetics and throughput. These results clearly demonstrate the potential of using Countercurrent Tangential Chromatography for low-cost commercial antibody purification.
 

12:30 Sponsored Luncheon Presentation (Opportunity Available) or Lunch on Your Own

 

ANALYTICAL STRATEGIES 

1:55 Chairperson’s Remarks

Yun (Kenneth) Kang, Ph.D., Principal Scientist & Head, Purification Team, Bioprocess Sciences, ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company

2:00 Gaining Efficiency and Value During Process Characterization

Rick St.JohnRick St. John, Ph.D., Senior Engineer, Purification Development, Biologics Department, Genentech, Inc.





 

2:30 Model-Based Rational Strategy for Chromatographic Resin Selection

Marcel OttensMarcel Ottens, Ph.D., Assistant Professor, BioSeparation Technology, Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology

In this presentation, a model‐based rational strategy for the selection of chromatographic resins is proposed and illustrated by evaluating three mixed mode adsorbents for the separation of a ternary model mixture of BSA, ovalbumin and amyloglucosidase. The main question addressed is selecting the most optimal chromatographic resin from a few promising alternatives. The methodology starts with chromatographic modeling, parameters acquisition and model validation, followed by model based optimization of the chromatographic separation for the resins of interest. Finally, the resins are rationally evaluated based on their optimized operating conditions and performance metrics such as product purity, yield, concentration, throughput, productivity and cost. The proposed model based approach could be a suitable alternative to column scouting during process development, the main strengths being the fact that resins are evaluated under their ideal working conditions, enabling a fair comparison. This presentation also demonstrates the application of column modeling and optimization to mixed mode chromatography.

3:00 Monitoring and Controlling Critical Purification Attributes of Viral Glycoproteins as Vaccine Product Candidates

Julie Q. Hang, Ph.D., Senior Scientist and Group Leader, Vaccine Protein Biochemistry, MedImmune LLC

Many recombinant viral surface glycoproteins are subunit vaccine targets in pre-clinical and clinical trials to induce humoral immune responses. Due to the complex nature of glycosylation, oligomerization and aggregation, purification products could introduce the product biophysical characteristics depending on purification processes. The changes showed impact on the vaccine immunogenicity. The strategies and case examples will be presented on how to use protein characterization tools to identify the critical purification attributes, and how to establish robust purification process to achieve consistent production specifications.

3:30 Refreshment Break with Exhibit and Poster Viewing

 

SPEEDING PURIFICATION PROCESSES 

4:15 Three-Phase Partitioning in Micro- and Macro-Scale Protein Purification

William Ward, Ph.D., Associate Professor, Biochemistry and Microbiology, Rutgers University - Biography 

Three-phase partitioning (TPP) was introduced in 1997, but this powerful protein purification technique has received only moderate recognition. Unlike any protein purification method known, TPP will semi-selectively release recombinant proteins from E. coli, purify the released protein to 80% homogeneity and remove all chromosomal DNA, scatter, turbidity, and viscosity. Simultaneously, TPP removes lipids, cell walls, and most pigments while concentrating the sample as much as 50-fold. All this is accomplished in less than 60 minutes and with minimal equipment. TPP is applicable to protein mini-preps and large scale purification processes.

4:45 The Protein Miniprep: High Purity Protein in 60 Minutes

David O’Connell, Ph.D., Senior Scientist, School of Medicine, University College Dublin - Biography 

In protein preparation, the purification of highly purified functional protein is rarely described as very quick and very easy. This presentation shows novel approaches to gently purify proteins very quickly in a highly simplified manner, accessible to the novice and expert alike. This approach has the potential to become a staple protocol for all sizes of laboratory, academic to industrial. This novel and powerful approach will be highlighted with interesting case studies of protein preparations of different functional proteins.

5:15 End of Conference


6:00 - 9:00 pm Recommended Dinner Short Course* 

SC6 Transient Protein Production in Mammalian Cells: A Short Story 


*Separate registration required 



 

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