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Affinity Protein Purification - Day 1


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Affinity Protein Purification

One of the more efficient methods to enrich or purify a protein from other proteins and components in a crude cell lysate or other samples is affinity purification, whereby the protein of interest is purified by taking advantage of its specific binding properties to an immobilized ligand. It is important to select the optimal method for your purification project. To maintain a competitive advantage, many issues need to be addressed such as fusion tags, support for affinity purification, quality and efficiency. This conference addresses key questions such as: Which resins are optimal for which procedure? Do tags need to be removed and if so, how? What are the newest tagging technologies? Is there an optimal way to ensure high yields, high speed and low costs?


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Monday, August 22

11:30 am Main Conference Registration

 

AFFINITY TAGS

1:00 pm Chairperson’s Opening Remarks

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

1:10 Featured Presentation

Affinity Tags, Their Uses and Removal

Paul Ramage, Ph.D., CPC, Protease Platform, Senior Research Investigator, Novartis Pharma AG

Affinity tags are widely used in protein science. Some are used to boost expression (both soluble and insoluble), some to increase solubility and many to specifically capture target proteins, especially when poorly expressed. In my talk I shall describe the tagging strategies we use for isolating recombinant proteases and related proteins, as well as the advantages and disadvantages of using such strategies.

1:45 Self-Cleaving Affinity Tags: A New Platform for Biologics Manufacturing?

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

The development of self-cleaving affinity tag technology has the potential to provide a new platform for biologics manufacturing, and several systems have been developed. The use of these technologies, however, will require an understanding of their impact on widely accepted downstream processes, as well as new validation methods for safe adoption and regulatory approval. This talk will examine these issues, with an emphasis on the specific opportunities for self-cleaving tags, as well as the remaining barriers to their application. This examination will include examples of how existing canonical processes might be impacted by the adoption of self-cleaving tag methods.

Sponsored by
Forte Bio 
2:15 The three A’s of Biologicals are Activity, Activity, Activity: Case Study of Guiding Process Development Towards Biological Specific Activity though Bioactivity and in Vitro Kinetic Assays

Oren Beske, Ph.D., Vice President, in Vitro Services, Aragen Biosciences
Monitoring quality and activity of a biological are essential during manufacturing development. This study describes significant bioactivity loss after adoption to an industrial CHO platform. Using the ForteBio Octet, an in vitro binding assay was rapidly developed, revealing a faster dissociation constant for the inactive form. Alternative clones and upstream processes were developed to increase specific activity.  Both the bioactivity assay and the ForteBio kinetic assay were used to track product quality during process development.

2:45 Networking Refreshment Break

3:15 Visible Heme Tags for Protein Affinity Purification and Quantification

Kara L. Bren, Ph.D., Professor, Chemistry, University of Rochester

A heme tag imparts intense color to a target protein for visual tracking during expression in E. coli and purification. Heme tags also can be utilized in affinity purification and protein quantification. Developments in heme tags will be discussed including achievement of tagging in the E. coli cytoplasm as well as periplasm.

3:45 High-Recovery Tandem Affinity Purification Tags

Yifeng Li, Ph.D., Technical Director, Protein Production Core Facility, Department of Biochemistry, The University of Texas Health Science Center

TAP coupled with mass spectrometry constitutes a powerful tool for the characterization of protein complex associated with a given target protein. This talk reviews several alternative TAP tags with improved efficiency and/or flexibility. In particular, a newly developed SBP-His tandem tag will be discussed in more detail.

4:15 Moderated Small-Group Breakout Discussions

Discussion groups give participants an opportunity to network and discuss important topics with colleagues from around the world.  We set aside this time for conference attendees to interact around a focused discussion topic, get to know one another, and develop contacts.  Meeting delegates select what discussion they would like to join from the list of topics provided, and the talk begins. The discussions are engaging, and a great way to network, exchange information, and establish future collaborations.

Topic: Native or Tagged--Getting a Handle on Proteins.

Moderator: Geoffrey S. Waldo, Ph.D., Team Leader, Biosciences, Los Alamos National Laboratory

When do you do native purifications and when do you use tags?
What do you use you tags for?
How do you get your tags off?
What are the pros and cons of tags?

Topic: Which Tagging Strategies Work?

Moderator: Paul Ramage, Ph.D., CPC, Protease Platform, Senior Research Investigator, Novartis Pharma AG

Topic: Self-Cleaving Tag Developments and Issues

Moderator: David Wood, Ph.D., Associate Professor, Chemical & Biomolecular Engineering,Ohio State University

5:30 Grand Opening Reception with Exhibit and Poster Viewing

7:00 End of Day One



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