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Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method

A new accurate spectrophotometric method for protein determination on nanoparticles is described. The method is based on the Coomassie blue dye that binds to the basic and aromatic amino acid residues of proteins, especially arginine and lysine. A known amount of reagent dye was mixed with a variety...

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Autores principales: Oviedo, Mariana J., Quester, Katrin, Hirata, Gustavo A., Vazquez-Duhalt, Rafael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6812394/
https://www.ncbi.nlm.nih.gov/pubmed/31667112
http://dx.doi.org/10.1016/j.mex.2019.09.015
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author Oviedo, Mariana J.
Quester, Katrin
Hirata, Gustavo A.
Vazquez-Duhalt, Rafael
author_facet Oviedo, Mariana J.
Quester, Katrin
Hirata, Gustavo A.
Vazquez-Duhalt, Rafael
author_sort Oviedo, Mariana J.
collection PubMed
description A new accurate spectrophotometric method for protein determination on nanoparticles is described. The method is based on the Coomassie blue dye that binds to the basic and aromatic amino acid residues of proteins, especially arginine and lysine. A known amount of reagent dye was mixed with a variety of protein-loaded nanoparticles. Thereafter the unconjugated reagent was mixed with excess protein (bovine serum albumin) and titrated. In this method, the reacted dye on the protein coating of nanoparticle is directly determined, in opposite to the conventional method, in which the conjugated protein is determined as the difference between the non-conjugated protein found in the supernatant after centrifugation, and the total amount of protein originally used. This method is able to measure amounts of coated protein lower than 1 ppm. • Simple and accurate method especially adapted for protein-coated nanoparticles. • The amino acid residues of protein in the nanoparticle surface react with Coomassie brilliant blue dye. • The unreacted dye is titrated with an excess of a standard protein.
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spelling pubmed-68123942019-10-30 Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method Oviedo, Mariana J. Quester, Katrin Hirata, Gustavo A. Vazquez-Duhalt, Rafael MethodsX Materials Science A new accurate spectrophotometric method for protein determination on nanoparticles is described. The method is based on the Coomassie blue dye that binds to the basic and aromatic amino acid residues of proteins, especially arginine and lysine. A known amount of reagent dye was mixed with a variety of protein-loaded nanoparticles. Thereafter the unconjugated reagent was mixed with excess protein (bovine serum albumin) and titrated. In this method, the reacted dye on the protein coating of nanoparticle is directly determined, in opposite to the conventional method, in which the conjugated protein is determined as the difference between the non-conjugated protein found in the supernatant after centrifugation, and the total amount of protein originally used. This method is able to measure amounts of coated protein lower than 1 ppm. • Simple and accurate method especially adapted for protein-coated nanoparticles. • The amino acid residues of protein in the nanoparticle surface react with Coomassie brilliant blue dye. • The unreacted dye is titrated with an excess of a standard protein. Elsevier 2019-09-14 /pmc/articles/PMC6812394/ /pubmed/31667112 http://dx.doi.org/10.1016/j.mex.2019.09.015 Text en © 2019 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Materials Science
Oviedo, Mariana J.
Quester, Katrin
Hirata, Gustavo A.
Vazquez-Duhalt, Rafael
Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method
title Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method
title_full Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method
title_fullStr Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method
title_full_unstemmed Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method
title_short Determination of conjugated protein on nanoparticles by an adaptation of the Coomassie blue dye method
title_sort determination of conjugated protein on nanoparticles by an adaptation of the coomassie blue dye method
topic Materials Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6812394/
https://www.ncbi.nlm.nih.gov/pubmed/31667112
http://dx.doi.org/10.1016/j.mex.2019.09.015
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