Wyatt SEC-MALS system

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Project Summary/Abstract Health care research has never been more critical than during a global crisis. At the well-established X-ray crystallography facility at The Pennsylvania State University (Penn State), we seek funds to replace our outdated Viscotek802 dynamic light scattering (DLS) equipment with a modern, Wyatt Technology DLS, multi angle light scattering (MALS) equipment in-line with size exclusion chromatography (SEC). This will enable accurate size, molecular weight, and stoichiometry data on a host of disease related biomolecules, viral and bacterial proteins, motor, nucleosome and intrinsically disordered proteins, nucleic acids, liposomes and their complexes. The Viscotek802 has been a workhorse for over fifteen years and served us well. Upgrading to a modern SEC-MALS capability, we will cater competently to the biophysical and structural characterization needs of our many exceptional NIH funded faculty and get the facility closer to an integrated structural biology approach. We have identified the Wyatt components as the sole commercial product which meets our requirements, is state-of-the-art and cost-effective. The Wyatt DLS and SEC- MALS system has a high accuracy, good resolution and superior sensitivity and is paramount for the success of the seventeen cutting edge health care NIH projects described in this proposal- (a) protein oligomers in the Hancock, Weinert, Yengo, Boehr, Cotruvo and Llinas labs (b) protein complexes in the Tan, Krasilnikov and Chroneos labs (c) RNA in the Bevilacqua lab (d) intrinsically disordered proteins in the Showalter and Zhang labs and (e) liposomes in the Tian lab. The ability to follow up the SEC-MALS run with an in-house small angle X-ray scattering (SAXS) run on the same sample is an exciting and novel configuration. The MALS-SAXS setup is designed aptly, with the output port of the Wyatt SEC-MALS system matching the input of the Rigaku-bioSAXS. With this handshake, we would be investing in true synergy where experimental results from SEC-MALS would guide the interpretation of SAXS and low-resolution solvent envelope models as obtained from SAXS could help in the crystallographic and CryoEM studies at higher resolution. The SEC-MALS instrument will be used by thirteen structural biology faculty from two Penn State campuses, University Park and Hershey, and three colleges, College of Medicine, Eberly College of Science, and the College of Engineering. No other functional SEC-MALS equipment exists in any lab on any of our twenty-four campuses. With a state-of-the-art Wyatt SEC-MALS system, our goal is to deduce accurate size, molecular weight, and stoichiometry in a wide spectrum of applications and enhance the capability at the Penn State University X-ray crystallography facility.