Protein Purification & Recovery
January 21-22, 2013
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TUESDAY, JANUARY 22
7:15 am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee
8:15 Chairperson’s Remarks
Suparna Mundodi, Ph.D., Global Product Manager, Global Marketing, Rainin Instrument, LLC
8:20 Sulfhydryl Oxidase-Dependent Expression of Recombinant Antibodies in Bacterial Cytoplasm
Ario de Marco, Ph.D., Director, Therapeutic Antibody Platform, Institut Curie – Paris - Biography
The expression of functional disulfide-bond dependent proteins in the reducing bacterial cytoplasm can be significantly improved by co-expressing the enzyme sulfhydryl oxidase. Specifically, we demonstrated that this approach is suitable for yielding recombinant antibodies that conserve their functionality. The opportunity of producing in vivo stable antibodies in the same cell compartment hosting their antigens allows for co-purification strategies, and using immunopurification for recovering difficult targets such as membrane proteins.
8:50 Development of a Non-Protein A Purification Platform for a Unique Next-Generation Antibody, the κλ-Body
Jean François Depoisier, Head, Downstream Processing, NovImmune - Biography
The κλ-Body has a molecular structure identical to a fully human monoclonal antibody. The talk will discuss the influence of this innovative bispecific antibody format on design of a robust non-protein A platform purification process. In order to exploit novel mechanisms of action and achieve superior clinical efficacy, NovImmune has developed a novel bispecific antibody format, the κλ-Body. The use of novel purification technologies will be discussed, notably involving new affinity ligands and mixed-mode chromatography.
9:20 Strategies for Aggregate Removal for a Non-Platform Antibody
Ruby Casareno, Ph.D., Principal Scientist, BioProcess Development, Seattle Genetics, Inc. - Biography
Aggregate removal was addressed at multiple points during the purification process. The unit operations adjusted to reduce aggregates were the Protein A capture step and filtration of viral inactivated pool. This antibody has high aggregate as a result of a high titer process. I’ll present alternative strategies to address aggregate removal based on process knowledge gained during initial development for this antibody as well as process experience gained from working with multiple antibodies in our pipeline.
9:50 Exploration of Proteomic Workflows in a Pipette Tip
Suparna Mundodi, Ph.D., Global Product Manager, Global Marketing, Rainin Instrument, LLC - Biography
The PureSpeed Protein Purification System carries out affinity chromatography using resin constrained within pipette tips, resulting in highly concentrated and purified protein. The PureSpeed System purifies recombinant proteins and antibodies, and functions in protein and chromatin immunoprecipitation, offering speed and versatility compared to other technologies.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing
10:45 The Influence of Histidine Tag Attachment on Protein Dynamics
Megan C. Thielges, Ph.D., Assistant Professor, Department of Chemistry, Indiana University - Biography
Significant changes appeared in the short time scale (picoseconds) dynamics of myoglobin as a result of His-tag incorporation in experiments employing two-dimensional infrared vibrational echo spectroscopy to measure the fluctuations of the CO vibrational frequency, which reports on protein structural dynamics. In this talk, I will introduce the methods of infrared spectroscopy for studying protein motion, describe our results with the protein myoglobin, and share our cautionary experiences with the impact of the His-tag incorporation.
11:15 Influence of Histidine-Tag on Structure and Immunological Potency of the Pre-Erythrocytic Stage Malaria Vaccine Candidate, P. falciparum CelTOS
Evelina Angov, Ph.D., Microbiologist, Military Malaria Research Program, Malaria Vaccine Branch, Walter Reed Army Institute of Research (WRAIR) - Biography
We previously reported on two relatively similar proteins that were impacted differently by the addition of the His-tag on their secondary structure and the induction of humoral and cellular immune responses. We expand on these findings by analyzing a third malaria protein, P. falciparum CelTOS. Preliminary findings suggest that in this case, the Histidine tag may have yielded a stabilizing effect on the molecule leading to the induction of ‘biologically’ relevant immune responses.
11:45 Close of Morning Session
12:30 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own
2:00 BuzZ Session A (Please visit our website for a listing of topics.)
2:45 Refreshment Break in the Exhibit Hall with Poster Awards
3:30 BuzZ Session B (Please visit our website for a listing of topics.)
4:15 End of Conference
Day 1| Day 2 | Download Pipeline 2 Brochure