Journal article
Temperature-Insensitive Near-Infrared Method for Determination of Protein Concentration during Protein Crystal Growth
Analytical chemistry (Washington), Vol.72(20), pp.4985-4990
10/01/2000
DOI: 10.1021/ac000406u
PMID: 11055719
Abstract
A temperature-insensitive method for measuring protein concentration in aqueous solutions using near-infrared spectroscopy is described. The method, which is based on identification of the net analyte signal of single-beam spectra, can be calibrated using a single protein absorbance measurement and is thus well suited for crystallization monitoring where the quantity of protein is limited. Then method is applied to measurements of glucose-isomerase concentration in a sodium phosphate buffer that is actively varied over the temperature range of 4-24 °C. The standard error of prediction using the optimized spectral range of 4670-4595 cm-1 is 0.12 mg/mL with no systematic trend in the residuals with solution temperature. The method is also applied to previously collected spectra of hen egg-white lysozyme and yields a standard error of prediction of 0.14 mg/mL. Spectra sampled at discrete wavelengths can also be used for calibration and prediction with performance comparable to that obtained with spectral bands. A set of four wavelengths are identified that can be used to predict concentrations of both proteins with a standard error less than 0.14 mg/mL.
Details
- Title: Subtitle
- Temperature-Insensitive Near-Infrared Method for Determination of Protein Concentration during Protein Crystal Growth
- Creators
- J. T Olesberg - Department of Chemistry and the Optical Science and Technology Center, The University of Iowa, Iowa City, Iowa 52242M. A Arnold - Department of Chemistry and the Optical Science and Technology Center, The University of Iowa, Iowa City, Iowa 52242Shih-Yao B Hu - Department of Chemistry and the Optical Science and Technology Center, The University of Iowa, Iowa City, Iowa 52242John M Wiencek - Department of Chemical and Biochemical Engineering and the Optical Science and Technology Center, The University of Iowa, Iowa City, Iowa 52242
- Resource Type
- Journal article
- Publication Details
- Analytical chemistry (Washington), Vol.72(20), pp.4985-4990
- DOI
- 10.1021/ac000406u
- PMID
- 11055719
- ISSN
- 0003-2700
- eISSN
- 1520-6882
- Language
- English
- Date published
- 10/01/2000
- Academic Unit
- Physics and Astronomy; Center for Biocatalysis and Bioprocessing; Fraternal Order of Eagles Diabetes Research Center; Chemistry
- Record Identifier
- 9984216577902771
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