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Protein Purification & Recovery header


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Conference Short Courses*View Details 

Sunday, January 9 - 3:00 pm - 6:00 pm

  • Protein Crystallization - Delineating Protein Structure
  • DoE and QbD: Tools for Optimizing the Bioprocess

Thursday, January 13 - 6:30 pm - 9:00 pm

  • Rational Design of Protein Solutions

 

This annual meeting provides an overview of the current tactics and technologies for optimizing the task of purifying proteins in the effort to reach consistency and quality. Case studies are presented that are based on science, yet illustrate the tenacity needed to maximize purity and yield, while minimizing purification steps. Sessions highlight alternative expression systems, affinity tags, achieving higher throughput, and difficult-to-purify proteins.

MONDAY, JANUARY 10


7:30am Conference Registration and Morning Coffee

 

Innovative Solutions

8:30 Chairperson’s Opening Remarks

William Gillette, Ph.D., Senior Scientist, Protein Expression Laboratory, SAIC-Frederick, Inc.

Opening Keynote Presentation:

8:45 Endo- and Exoproteolytic Removal of Affinity Tags

David WaughDavid S. Waugh, Ph.D., Chief, Protein Engineering Section, Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick

Methods and reagents for the removal of affinity tags will be described, with emphasis on the use of tobacco etch virus (TEV) protease and carboxypeptidases.  In particular, recent advances in the production and biochemical characterization of an A-type carboxypeptidase from Metarhizium anisopleae will be presented.



Featured Presentation:

9:35 New Insights into Parvovirus Filtration for Monoclonal Antibody Purification

Amit Mehta, Ph.D., Senior Engineer & Group Leader, Genentech Inc.

 

10:05 Networking Coffee Break

 

Optimizing Purification Processes

10:45 Optimizing the Recovery of Protein Targets Using Statistical Design of Experiment

Susanne Swalley, Ph.D., Research Scientist II, Vertex Pharmaceuticals, Inc.

Advances in high-throughput protein expression and purification have made it possible to apply statistical design of experiment (DoE) to the field of protein sciences. By varying numerous factors in parallel we can rapidly optimize protein yield and purity. Additionally, we can take advantage of statistical data analysis to visualize interactions among multiple experimental variables, allowing for reliable prediction in areas not directly covered by experimentation. We will describe the application of DoE to the preparation of both cytosolic and membrane proteins for functional and structural studies.

11:15 Evaluation of a Multimodal Resin for Selective Capture of CHO-Derived Monoclonal Antibodies Directly from Harvested Cell Culture Fluid

Kimberly A. Kaleas, Engineer II, BioProcess Development, Pharma Technical Development, Genentech, a Member of the Roche Group

Multimodal chromatography resins are gaining popularity as effective purification tools. This proof-of-concept study examines Capto MMC resin as a capture step for monoclonal antibody (mAb) purification from harvested cell culture fluid. Binding properties of >10 CHO-derived mAb feedstocks were examined for dynamic binding capacity and 3 separate elution strategies were identified. Two feedstocks were used to further explore the purification potential of this technology.

11:45 Solutions for the Expression and Purification of Pre-Clinical Grade Therapeutic Proteins

Richard J. Buick, Ph.D., Technical Director, Fusion Antibodies, Ltd.

Recombinant proteins and antibodies are regularly used for preclinical evaluation in in vivo models.  We present case studies of expression and purification strategies to prepare up to gram levels of protein or antibody in a non-cGMP facility, with high purity and low contaminant levels suitable for preclinical studies.  The expression has been performed in both bacterial and mammalian systems.

Sponsored by
BAC
12:15pm Filling up the Gaps in Antibody Purification
Pim Hermans, Director of Ligand Discovery, R&D, BAC b.v.
Since more and more antibody based therapeutics are lacking a regular protein A binding site it becomes a challenge to find a protein A equivalent for primary capture. For this reason BAC has developed a range of CaptureSelect affinity ligands directed against a unique panel of antibody sub-domains (e.g. present on Fab) to facilitate affinity purification of virtually any antibody based format.

 

Sponsored by
PromegaNEW
 12:30 Luncheon Presentation
Robust Purification of Arginine Methyltransferase (CARM1) from Mammalian Cells Using HaloTag Technology

Rob Chumanov, Ph.D., Promega
Identification and characterization of post-translational modifications and protein function largely depends on success in purification of full-length target protein. CARM1 is a mammalian transcription factor that plays a role in progression of breast cancer. CARM1 is able to methylate itself; however, this process has been previously uncharacterized, due to difficulty in purifying the full-length protein. HaloTag technology enabled the purification of full-length, active CARM1 from mammalian cells; the purified protein was used for both mass spectrometry and functional analysis to characterize the site of automethylation.  This work represents the first identified physiological role for automethylation in the protein arginine methyltransferase family.



1:00 Sponsored Luncheon Presentation (Opportunity Avaialble) or Lunch on your Own 

 

Tag Technology

2:00 Chairperson’s Remarks

Alexey Veraksa, Ph.D., Assistant Professor, Biology, College of Science & Mathematics, University of Massachusetts, Boston

2:05 Self-Cleaving Tags: New Options and New Applications

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

Self-cleaving affinity and non-chromatographic tags have substantial potential for impacting the manufacture of biologics. This presentation will summarize recent results in demonstrating these methods with industrially relevant products and processes, as well as compare newer systems for generating self-cleaving tags. Ultimately, these technologies can provide general platforms for rapid process and product development, while simultaneously decreasing costs associated with downstream processing.

2:35 The Biotinylated Sortase Self-Cleavage Purification Method (BISOP) Combined with the Wheat Germ Cell-Free System

Yaeta Endo, Ph.D., Professor & Director, Cell-Free Science and Technology Research Center, Ehime University, Matsuyama, Japan

Among cell-free protein production systems, the one we developed using wheat germ is of special interest for its eukaryotic nature; it has significant advantages in the parallel production of a large number of eukaryotic multi-domain proteins in folded state. In this presentation we will focus on introducing the BISOP method, a new protein purification method based on the wheat germ cell-free system and describe its use to prepare tag-free, high quality proteins, for functional and structural analyses.

3:05 PEGylate and Purify: A Dual Role of His-tag for PEGylated Therapeutics

Ji-won Choi, Ph.D., Director, Biology, PolyTherics, Ltd.

Genetically fused his-tags are widely used to facilitate expression and purification of proteins with therapeutic potential. The properties of the next generation therapeutic protein products, known as biobetters, is enhanced by technologies such as PEGylation, which increases the duration of action of drugs by increasing their in vivo half-life through reducing their rate of clearance. We have harnessed the advantages of both technologies by developing a unique technology, known as HiPEGTM, that conjugates PEG to polyhistidine sequences and then shown that it is possible to purify the PEG-conjugated protein by immobilised metal affinity chromatography (IMAC) using the same his-tag.


Sponsored by
Lab901

3:35 At-line SDS-PAGE for Biologic Production: Insights into how Process Parameters
Affect mAb Yield
Adam Inche, Product Manager, Protein Products, Lab901 Ltd
Conventional SDS-PAGE can be too time consuming and costly to allow at-line production monitoring of bio-pharmaceuticals. ScreenTape is a unique solution to this issue which not only delivers SDS-PAGE results in as little as 1 minute per sample, but unused lanes are available for subsequent analyses. Information on product quantity and quality is delivered automatically in an electronic format for easy data manipulation and archiving. This talk will show how ScreenTape can be employed to monitor mAb production at-line by SDS-PAGE, and deliver unprecedented information on how different process parameters can affect product yield.



3:50 Networking Refreshment Break 
 

Difficult to Purify

4:30 Using Tandem Affinity Purification-Mass Spectrometry for Analyzing Intracellular Signaling Complexes

Alexey Veraksa, Ph.D., Assistant Professor, Biology, College of Science & Mathematics, University of Massachusetts, Boston

I will discuss the advantages and pitfalls associated with the use of the tandem affinity purification (TAP)-mass spectrometry, and will share our new data on developing even more efficient ways to isolate and analyze protein complexes. The Notch signaling pathway is a conserved mechanism of intercellular communication that is used in all metazoans to establish molecular differences between adjacent cells. We have used the original TAP tag, as well as more recently developed GS-TAP tag, to analyze protein interactions in the Notch pathway.

5:00 Purification and Stabilization of Membrane Proteins

Lawrence DeLucas, Ph.D., Director, Center for Biophysical Sciences & Engineering, University of Alabama, Birmingham

In the past two years we have demonstrated success establishing critical underpinnings for protein expression in mammalian cells. This expression system is combined with an early assessment purification protocol that reduces the time and costs associated with determination of the optimum purification strategy. Finally, our approach incorporates a novel high-throughput technology, applicable to aqueous and membrane proteins, that can improve protein solubility, physical stability and crystallization success.

5:30 Development of a Manufacturable, Cost Effective Nickel Affinity Purification for the Purification of Insoluble Recombinant Proteins

Troy McSherry, Ph.D., Principal Scientist, Diagnostics Division, Abbott Laboratories

Purification process development today requires an eye on process cost as well as technical approach consideration. We have developed a nickel affinity purification process for the purification of insoluble, tagged recombinant proteins. Taking advantage of the necessary denaturation of the protein required for purification, we were able to obtain high binding capacities and purities compared to purification of soluble native proteins. This initial proof of concept work was just the beginning however, and was followed by significant development and characterization work aimed at maximizing process manufacturability, reproducibility, robustness, high yield and overall cost effectiveness.

6:00-7:30 Welcoming Reception in the Exhibit Hall

 

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Links to Companion Meetings

Pipeline 1

Protein Data Integration and Interrogation 

Protein Computational Tools