A Decoupled Automation Platform for Hydrogen/Deuterium Exchange Mass Spectrometry Experiments.

Autor: Espada A; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Haro R; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Castañon J; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Sayago C; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Perez-Cozar F; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Cano L; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Redero P; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Molina-Martin M; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Broughton H; Centro de Investigación Lilly, SA, Avenida de la Industria 30, 28108, Alcobendas, Spain., Stites RE; Eli Lilly and Company, Lilly Corporate Center, Lilly Research Laboratories, Indianapolis, IN, 46285, USA., Pascal BD; Omics Informatics LLC, 1050 Bishop Street #517, Honolulu, HI, 96813, USA., Griffin PR; Department of Molecular Medicine, The Scripps Research Institute, 130 Scripps Way, Jupiter, FL, 33458, USA., Dodge JA; Eli Lilly and Company, Lilly Corporate Center, Lilly Research Laboratories, Indianapolis, IN, 46285, USA., Chalmers MJ; Eli Lilly and Company, Lilly Corporate Center, Lilly Research Laboratories, Indianapolis, IN, 46285, USA. chalmers_michael@lilly.com.
Jazyk: angličtina
Zdroj: Journal of the American Society for Mass Spectrometry [J Am Soc Mass Spectrom] 2019 Dec; Vol. 30 (12), pp. 2580-2583. Date of Electronic Publication: 2019 Nov 13.
DOI: 10.1007/s13361-019-02331-2
Abstrakt: Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a biophysical technique well suited to the characterization of protein dynamics and protein-ligand interactions. In order to accurately define the rate of exchange, HDX experiments require the repeated measure of deuterium incorporation into the target protein across a range of time points. Accordingly, the HDX-MS experiment is well suited to automation, and a number of automated systems for HDX-MS have been developed. The most widely utilized platforms all operate an integrated design, where robotic liquid handling is interfaced directly with a mass spectrometer. With integrated designs, the exchange samples are prepared and injected into the LC-MS following a "real-time" serial workflow. Here we describe a new HDX-MS platform that is comprised of two complementary pieces of automation that disconnect the sample preparation from the LC-MS analysis. For preparation, a plate-based automation system is used to prepare samples in parallel, followed by immediate freezing and storage. A second piece of automation has been constructed to perform the thawing and LC-MS analysis of frozen samples in a serial mode and has been optimized to maximize the duty cycle of the mass spectrometer. The decoupled configuration described here reduces experiment time, significantly improves capacity, and improves the flexibility of the platform when compared with a fully integrated system.
Databáze: MEDLINE
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