Attenuated total reflection-FT-IR spectroscopic imaging of protein crystallization

K. L. Andrew Chan, Lata Govada, Roslyn M. Bill, Naomi E. Chayen, Sergei G. Kazarian

Research output: Contribution to journalArticlepeer-review

Abstract

Protein crystallization is of strategic and commercial relevance in the post-genomic era because of its pivotal role in structural proteomics projects. Although protein structures are crucial for understanding the function of proteins and to the success of rational drug design and other biotechnology applications, obtaining high quality crystals is a major bottleneck to progress. The major means of obtaining crystals is by massive-scale screening of a target protein solution with numerous crystallizing agents. However, when crystals appear in these screens, one cannot easily know if they are crystals of protein, salt, or any other molecule that happens to be present in the trials. We present here a method based on Attenuated Total Reflection (ATR)-FT-IR imaging that reliably identifies protein crystals through a combination of chemical specificity and the visualizing capability of this approach, thus solving a major hurdle in protein crystallization. ATR-FT-IR imaging was successfully applied to study the crystallization of thaumatin and lysozyme in a high-throughput manner, simultaneously from six different solutions. This approach is fast as it studies protein crystallization in situ and provides an opportunity to examine many different samples under a range of conditions.
Original languageEnglish
Pages (from-to)3769-3775
Number of pages7
JournalAnalytical Chemistry
Volume81
Issue number10
DOIs
Publication statusPublished - 23 May 2009

Bibliographical note

© 2009 American Chemical Society. This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://dx.doi.org/10.1021/ac900455y

Keywords

  • protein crystallization
  • post-genomic era
  • structural proteomics projects
  • rational drug design

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