Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensing
Introduction: Gold nanoparticles (AuNPs) and their conjugates are used for many applications in the field of sensors. Literature lacks procedures able to separate, purify and characterize these species in native conditions without altering them while assuring a high throughput. This technological ga...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-12-01
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| Series: | Frontiers in Sensors |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fsens.2022.1087115/full |
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| author | Junjie Wang Stefano Giordani Valentina Marassi Valentina Marassi Barbara Roda Barbara Roda Pierluigi Reschiglian Pierluigi Reschiglian Andrea Zattoni Andrea Zattoni |
| author_facet | Junjie Wang Stefano Giordani Valentina Marassi Valentina Marassi Barbara Roda Barbara Roda Pierluigi Reschiglian Pierluigi Reschiglian Andrea Zattoni Andrea Zattoni |
| author_sort | Junjie Wang |
| collection | DOAJ |
| description | Introduction: Gold nanoparticles (AuNPs) and their conjugates are used for many applications in the field of sensors. Literature lacks procedures able to separate, purify and characterize these species in native conditions without altering them while assuring a high throughput. This technological gap can be reduced by exploiting Asymmetrical Flow Field Flow Fractionation multidetection platforms (AF4 multidetection).Method: This work describes a complete set of strategies based on the AF4 system, from nanoparticle synthesis to separative method optimization to conjugates screening and characterization, achieving quantitative control and purification of ready-to-use conjugated Gold nanoparticles and ensuring effectiveness in biosensing.Results and Discussion: AF4-multidetection was used to study AuNPs with different types of surface coating [Poly ethylene glycol, (PEG) and Citrate], their binding behaviour with protein (Bovine serum albumin, BSA) and their stability after conjugation to BSA. A robust but flexible method was developed, able to be applied to different AuNPs and conjugating molecules. The morphology and conjugation mechanism of AuNPs-BSA conjugates were evaluated by combining online Multiangle light scattering (MALS) and offline Dynamic Light Scattering (DLS) measures, which provided an important feature for the quality control required to optimize bio-probe synthesis and subsequent bioassays. |
| first_indexed | 2024-04-13T06:01:11Z |
| format | Article |
| id | doaj.art-57a81264924942039530d0d26b90af62 |
| institution | Directory Open Access Journal |
| issn | 2673-5067 |
| language | English |
| last_indexed | 2024-04-13T06:01:11Z |
| publishDate | 2022-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Sensors |
| spelling | doaj.art-57a81264924942039530d0d26b90af622022-12-22T02:59:26ZengFrontiers Media S.A.Frontiers in Sensors2673-50672022-12-01310.3389/fsens.2022.10871151087115Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensingJunjie Wang0Stefano Giordani1Valentina Marassi2Valentina Marassi3Barbara Roda4Barbara Roda5Pierluigi Reschiglian6Pierluigi Reschiglian7Andrea Zattoni8Andrea Zattoni9Department of Chemistry G. Ciamician, University of Bologna, Bologna, ItalyDepartment of Chemistry G. Ciamician, University of Bologna, Bologna, ItalyDepartment of Chemistry G. Ciamician, University of Bologna, Bologna, ItalyByFlow SRL, Bologna, ItalyDepartment of Chemistry G. Ciamician, University of Bologna, Bologna, ItalyByFlow SRL, Bologna, ItalyDepartment of Chemistry G. Ciamician, University of Bologna, Bologna, ItalyByFlow SRL, Bologna, ItalyDepartment of Chemistry G. Ciamician, University of Bologna, Bologna, ItalyByFlow SRL, Bologna, ItalyIntroduction: Gold nanoparticles (AuNPs) and their conjugates are used for many applications in the field of sensors. Literature lacks procedures able to separate, purify and characterize these species in native conditions without altering them while assuring a high throughput. This technological gap can be reduced by exploiting Asymmetrical Flow Field Flow Fractionation multidetection platforms (AF4 multidetection).Method: This work describes a complete set of strategies based on the AF4 system, from nanoparticle synthesis to separative method optimization to conjugates screening and characterization, achieving quantitative control and purification of ready-to-use conjugated Gold nanoparticles and ensuring effectiveness in biosensing.Results and Discussion: AF4-multidetection was used to study AuNPs with different types of surface coating [Poly ethylene glycol, (PEG) and Citrate], their binding behaviour with protein (Bovine serum albumin, BSA) and their stability after conjugation to BSA. A robust but flexible method was developed, able to be applied to different AuNPs and conjugating molecules. The morphology and conjugation mechanism of AuNPs-BSA conjugates were evaluated by combining online Multiangle light scattering (MALS) and offline Dynamic Light Scattering (DLS) measures, which provided an important feature for the quality control required to optimize bio-probe synthesis and subsequent bioassays.https://www.frontiersin.org/articles/10.3389/fsens.2022.1087115/fullbiosensor developmentAuNPsnative separationFFF-multidetectionprotein conjugation |
| spellingShingle | Junjie Wang Stefano Giordani Valentina Marassi Valentina Marassi Barbara Roda Barbara Roda Pierluigi Reschiglian Pierluigi Reschiglian Andrea Zattoni Andrea Zattoni Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensing Frontiers in Sensors biosensor development AuNPs native separation FFF-multidetection protein conjugation |
| title | Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensing |
| title_full | Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensing |
| title_fullStr | Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensing |
| title_full_unstemmed | Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensing |
| title_short | Quality control and purification of ready-to-use conjugated gold nanoparticles to ensure effectiveness in biosensing |
| title_sort | quality control and purification of ready to use conjugated gold nanoparticles to ensure effectiveness in biosensing |
| topic | biosensor development AuNPs native separation FFF-multidetection protein conjugation |
| url | https://www.frontiersin.org/articles/10.3389/fsens.2022.1087115/full |
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