Article Dans Une Revue Analytical Biochemistry Année : 2015

Quantification of amyloid fibrils using size exclusion chromatography coupled with online fluorescence and ultraviolet detection

Résumé

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.

Dates et versions

hal-04155880 , version 1 (07-07-2023)

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Irina Randrianjatovo-Gbalou, Claire-Emmanuelle Marcato-Romain, Elisabeth Girbal-Neuhauser. Quantification of amyloid fibrils using size exclusion chromatography coupled with online fluorescence and ultraviolet detection. Analytical Biochemistry, 2015, 488, pp.19-21. ⟨10.1016/j.ab.2015.07.014⟩. ⟨hal-04155880⟩
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