Skip to content Skip to navigation
University of Warwick
  • Study
  • |
  • Research
  • |
  • Business
  • |
  • Alumni
  • |
  • News
  • |
  • About

University of Warwick
Publications service & WRAP

Highlight your research

  • WRAP
    • Home
    • Search WRAP
    • Browse by Warwick Author
    • Browse WRAP by Year
    • Browse WRAP by Subject
    • Browse WRAP by Department
    • Browse WRAP by Funder
    • Browse Theses by Department
  • Publications Service
    • Home
    • Search Publications Service
    • Browse by Warwick Author
    • Browse Publications service by Year
    • Browse Publications service by Subject
    • Browse Publications service by Department
    • Browse Publications service by Funder
  • Help & Advice
University of Warwick

The Library

  • Login
  • Admin

Infrared absorbance spectroscopy of aqueous proteins : comparison of transmission and ATR data collection and analysis for secondary structure fitting

Tools
- Tools
+ Tools

Corujo, Marco Pinto, Sklepari, Meropi, Ang, Dale L., Millichip, Mark, Reason, Andrew, Goodchild, Sophia C., Wormell, Paul, Amarasinghe, Don Praveen, Lindo, Viv, Chmel, Nikola Paul and Rodger, Alison (2018) Infrared absorbance spectroscopy of aqueous proteins : comparison of transmission and ATR data collection and analysis for secondary structure fitting. Chirality, 30 (8). pp. 957-965. doi:10.1002/chir.23002 ISSN 0899-0042.

[img]
Preview
PDF
WRAP-infrared-absorbance-spectroscopy-aqueous-proteins-analysis-Corujo-2018.pdf - Published Version - Requires a PDF viewer.
Available under License Creative Commons Attribution 4.0.

Download (658Kb) | Preview
Official URL: https://doi.org/10.1002/chir.23002

Request Changes to record.

Abstract

Attenuated total reflectance (ATR) infrared absorbance spectroscopy of proteins in aqueous solution is much easier to perform than transmission spectroscopy, where short path‐length cells need to be assembled reproducibly. However, the shape of the resulting ATR infrared spectrum varies with the refractive index of the sample and the instrument configuration. Refractive index in turn depends on the absorbance of the sample. In this work, it is shown that a room temperature triglycine sulfate detector and a ZnSe ATR unit can be used to collect reproducible spectra of proteins. A simple method for transforming the protein ATR spectrum into the shape of the transmission spectrum is also given, which proceeds by approximating a Kramers‐Krönig–determined refractive index of water as a sum of four linear components across the amide I and II regions. The light intensity at the crystal surface (with 45° incidence) and its rate of decay away from the surface is determined as a function of the wave number–dependent refractive index as well as the decay of the evanescent wave from the surface. The result is a single correction factor at each wave number. The spectra were normalized to a maximum of 1 between 1600 cm−1 and 1700 cm−1 and a self‐organizing map secondary structure fitting algorithm, SOMSpec, applied using the BioTools reference set. The resulting secondary structure estimates are encouraging for the future of ATR spectroscopy for biopharmaceutical characterization and quality control applications.

Item Type: Journal Article
Subjects: Q Science > QD Chemistry
Divisions: Faculty of Science, Engineering and Medicine > Science > Chemistry
Faculty of Science, Engineering and Medicine > Research Centres > Molecular Organisation and Assembly in Cells (MOAC)
SWORD Depositor: Library Publications Router
Library of Congress Subject Headings (LCSH): Absorbance scale (Spectroscopy), Absorption spectra, Proteins
Journal or Publication Title: Chirality
Publisher: Wiley
ISSN: 0899-0042
Official Date: August 2018
Dates:
DateEvent
August 2018Published
20 July 2018Available
14 February 2018Accepted
Volume: 30
Number: 8
Page Range: pp. 957-965
DOI: 10.1002/chir.23002
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access (Creative Commons)
Date of first compliant deposit: 21 September 2018
Date of first compliant Open Access: 21 September 2018
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
EP/K007394/1[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266
EP/F500378/1[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266
EP/L015307/1[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266
BB/F011199/1 [BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268

Request changes or add full text files to a record

Repository staff actions (login required)

View Item View Item

Downloads

Downloads per month over past year

View more statistics

twitter

Email us: wrap@warwick.ac.uk
Contact Details
About Us