Systematic process evaluation of the conjugation of proteins to gold nanoparticles
The present work addresses some fundamental aspects in the preparation of protein-conjugated gold nanoparticles, in order to ensure an appropriate final product. Ten broadly available and/or easy to implement analytical tools were benchmarked and compared in their capacity to provide reliable and co...
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doaj-545aa7e7741f4f4aad4c25b8ee6af6512021-07-05T16:35:01ZengElsevierHeliyon2405-84402021-06-0176e07392Systematic process evaluation of the conjugation of proteins to gold nanoparticlesPablo Fagúndez0Santiago Botasini1Juan Pablo Tosar2Eduardo Méndez3Unidad de Bioquímica Analítica, Centro de Investigaciones Nucleares, Facultad de Ciencias, Universidad de la República, 11400, Montevideo, Uruguay; Graduate Program in Chemistry, Facultad de Química, Universidad de la República, UruguayLaboratorio de Biomateriales, Instituto de Química Biológica, Facultad de Ciencias, Universidad de la República, 11400, Montevideo, UruguayUnidad de Bioquímica Analítica, Centro de Investigaciones Nucleares, Facultad de Ciencias, Universidad de la República, 11400, Montevideo, UruguayLaboratorio de Biomateriales, Instituto de Química Biológica, Facultad de Ciencias, Universidad de la República, 11400, Montevideo, Uruguay; Corresponding author.The present work addresses some fundamental aspects in the preparation of protein-conjugated gold nanoparticles, in order to ensure an appropriate final product. Ten broadly available and/or easy to implement analytical tools were benchmarked and compared in their capacity to provide reliable and conclusive information for each step of the procedure. These techniques included transmission electron microscopy, UV/VIS spectroscopy, dynamic light scattering, zeta-potential, Fourier-transformed infrared spectroscopy, colloidal stability titration, end-point colloidal stability analysis, cyclic voltammetry, agarose gel electrophoresis and size-exclusion chromatography (SEC). Four different proteins widely used as adaptors or blocking agents were tested, together with 13 nm gold nanoparticles containing different surface chemistries. Among all tested techniques, some of the least popular among nanomaterial scientists probed to be the most informative, including colloidal stability, gel electrophoresis and SEC; the latter being also an efficient purification procedure. These three techniques provide low-cost, low time consuming, sensitive and robust ways to assess the success of the nanoparticle bioconjugation steps, especially when used in adequate combinations.http://www.sciencedirect.com/science/article/pii/S240584402101495XBioconjugationColloidal stabilityGold nanoparticlesCovalent attachmentNon-covalent attachment |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Pablo Fagúndez Santiago Botasini Juan Pablo Tosar Eduardo Méndez |
spellingShingle |
Pablo Fagúndez Santiago Botasini Juan Pablo Tosar Eduardo Méndez Systematic process evaluation of the conjugation of proteins to gold nanoparticles Heliyon Bioconjugation Colloidal stability Gold nanoparticles Covalent attachment Non-covalent attachment |
author_facet |
Pablo Fagúndez Santiago Botasini Juan Pablo Tosar Eduardo Méndez |
author_sort |
Pablo Fagúndez |
title |
Systematic process evaluation of the conjugation of proteins to gold nanoparticles |
title_short |
Systematic process evaluation of the conjugation of proteins to gold nanoparticles |
title_full |
Systematic process evaluation of the conjugation of proteins to gold nanoparticles |
title_fullStr |
Systematic process evaluation of the conjugation of proteins to gold nanoparticles |
title_full_unstemmed |
Systematic process evaluation of the conjugation of proteins to gold nanoparticles |
title_sort |
systematic process evaluation of the conjugation of proteins to gold nanoparticles |
publisher |
Elsevier |
series |
Heliyon |
issn |
2405-8440 |
publishDate |
2021-06-01 |
description |
The present work addresses some fundamental aspects in the preparation of protein-conjugated gold nanoparticles, in order to ensure an appropriate final product. Ten broadly available and/or easy to implement analytical tools were benchmarked and compared in their capacity to provide reliable and conclusive information for each step of the procedure. These techniques included transmission electron microscopy, UV/VIS spectroscopy, dynamic light scattering, zeta-potential, Fourier-transformed infrared spectroscopy, colloidal stability titration, end-point colloidal stability analysis, cyclic voltammetry, agarose gel electrophoresis and size-exclusion chromatography (SEC). Four different proteins widely used as adaptors or blocking agents were tested, together with 13 nm gold nanoparticles containing different surface chemistries. Among all tested techniques, some of the least popular among nanomaterial scientists probed to be the most informative, including colloidal stability, gel electrophoresis and SEC; the latter being also an efficient purification procedure. These three techniques provide low-cost, low time consuming, sensitive and robust ways to assess the success of the nanoparticle bioconjugation steps, especially when used in adequate combinations. |
topic |
Bioconjugation Colloidal stability Gold nanoparticles Covalent attachment Non-covalent attachment |
url |
http://www.sciencedirect.com/science/article/pii/S240584402101495X |
work_keys_str_mv |
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