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Start Over Author "Girbal-Neuhauser E" Journal analytical biochemistry
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2. Quantification of amyloid fibrils using size exclusion chromatography coupled with online fluorescence and ultraviolet detection.

Author

Randrianjatovo-Gbalou I, Marcato-Romain CE, and Girbal-Neuhauser E

Subjects
Algorithms, Amyloid chemistry, Animals, Bacillus chemistry, Bacillus physiology, Bacterial Proteins analysis, Bacterial Proteins chemistry, Bacterial Proteins isolation & purification, Benzothiazoles, Biofilms, Calibration, Caseins analysis, Caseins chemistry, Cattle, Chromatography, Gel, Fluorescent Dyes chemistry, France, Molecular Weight, Protein Aggregates, Solubility, Spectrometry, Fluorescence, Spectrophotometry, Ultraviolet, Thiazoles chemistry, Amyloid analysis, Models, Molecular, Protein Aggregation, Pathological diagnosis
Abstract

An amyloid fibrils investigation within biofilm samples requires distinguishing the amyloid β-sheet structure of these proteins and quantifying them. In this study, the property of amyloids to incorporate the fluorescent dye Thioflavin T has been exploited to propose a method of quantification. The experimental protocol includes the preparation of amyloids from commercial κ-casein (κCN) and their fractionation through size exclusion chromatography (SEC) to provide calibration curves from fluorescence and absorbance signals. Finally, a bacterial biofilm extract was injected into SEC, and the amyloid fibrils could be expressed as equivalent κCN, representing approximately 21% of the total proteins., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Published
2015
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