Barnett Institute Retreat The Institute retreat this year emphasized and strengthened our interactions with affiliated and collaborating laboratories, including presentations by four colleagues.
List of Speakers
William Hancock, Bradstreet Chair
in Bioanalytical Chemistry, Barnett Inst.
Paul Vouros, Sr. Faculty Fellow,
Barnett Institute. Zhaohui 'Sunny' Zhou, Faculty
Fellow, Barnett Institute. Rohin Mhatre, Director, Bio Process
Development, Biogen Idec John Engen, Faculty Fellow, Barnett
Inst. Igor Kaltashov, Assoc. Prof. of
Bioanalytical Chemistry, UMASS Amherst
Summaries of Guest Lectures Hanno Steen is Director of the Core Facility for Proteomics at Children's Hospital in Boston, and is Faculty of Harvard Medical School. In The Cell Cycle Meets Proteomics, Hanno presented work mapping the protein phosphorylation patterns (activation events) of key proteins that regulate cell division. A surprising highlight was that 3 different anticancer drugs, which arrest cells in prometaphase by action on microtubules, and are thought to have a similar mode of action, produce markedly different phosphorylation patterns of the anaphase promoting complex (APC). Hanno's work was a vivid illustration of how analytical work is planned and interpreted in the context of the biological system under study. Rohin Mhatre Director of Bio Process Development at Biogen Idec spoke about The "Quality by Design" Initiative in the Biotechnology Industry. "Students need to be aware of this important initiative in industry". "Design Space", in biopharmaceutical production, is when you can evaluate the impact of the process inputs on the process outputs. To reach "design space" you need to know a lot about your molecule, from clinical and preclinical data, SAR and stability -- all of the critical quality attributes (CQA). You then correlate the critical attributes with detailed analysis of the molecule, and find ways to measure the important ones in production: Process Analytical Technology (PAT). For example, phosphorylation and glycosylation are a major concern. With a fast map using SIM for specific transitions, we can detect sialyation in 5 minutes, follow it in real time. Joe Zaia Assoc. Research Prof. of Biochemistry & Assoc. Dir., Mass Spectrometry Resource, Boston University School of Medicine An LC-MS Platform for Tissue-Based Glycomics. A large number of membrane-bound proteins are so heavily modified with glycosaminoglycans (GAG) that their primary structure resembles a bottle-brush. The GAG's in turn are highly heterogeneous, primarily repeating units of N-acetylglycosamine with hexuronic acid, glucuronic acid or a hexose. In some regions the NAG is heavily sulfated. Glycans are challenging to study; GAG's especially so. Dr. Zaia reviewed 9 LC methods for GAG analysis, favoring HILIC in general, and a chip-based separation and analysis device in particular. QTOF mass accuracy is limited for glycans, but an orbitrap gives 2 ppm, like peptides. In a comparisoin of several heparins, we saw the 6-sulfate and 7-sulfate forms bound to antithrombin III, the 8- and 9-sulfate forms had little affinity. A variety of low molecular weight heparins are on the market, each for a narrow clinical range. As patents expire, showing equivalency of generic products will be difficult. This MS assay may be able to define the product for QC.
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Hanno Steen, Director of the
Proteomics Center, Children’s Hospital, Boston
Joe Zaia, Assoc. Research Prof. of
Biochemistry & Assoc. Dir., Mass Spectrometry Resource, Boston
University School of Medicine
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