Cargando…

Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry

Tumor-derived exosomes may provide biomarkers for cancer treatment. Using sputtering technology, an affinity-based device to capture exosomes was developed using nanoporous substrate (NPG)-coated silicon microscopy. Immunology-based techniques detect and purify exosomes using gold coating with a spe...

Descripción completa

Detalles Bibliográficos
Autores principales: Al Mannai, Amera, Al-Ansari, Tareq, Saoud, Khaled M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9460504/
https://www.ncbi.nlm.nih.gov/pubmed/36080806
http://dx.doi.org/10.3390/s22176347
_version_ 1784786763760271360
author Al Mannai, Amera
Al-Ansari, Tareq
Saoud, Khaled M.
author_facet Al Mannai, Amera
Al-Ansari, Tareq
Saoud, Khaled M.
author_sort Al Mannai, Amera
collection PubMed
description Tumor-derived exosomes may provide biomarkers for cancer treatment. Using sputtering technology, an affinity-based device to capture exosomes was developed using nanoporous substrate (NPG)-coated silicon microscopy. Immunology-based techniques detect and purify exosomes using gold coating with a specific antigen. Inverted fluorescent microscopy was used to detect target exosomes quantitatively utilizing fluorescent nanospheres as the label. We quantified the expression of CD63 surface protein markers on exosomes from conditioned culture media of breast cancer cells. The exosomes that targeted specific proteins with controls were statistically analyzed and compared to those that targeted non-specific proteins. Results from SEM showed that the exosomes were circular, between 30 and 150 nanometers in size. The porous gold substrates captured more exosomes than the nonporous substrates. Nitric acid treatments at different times resulted in a variety of pore sizes. Despite the increase in the size of the pores, the number of exosomes found in the porous gold substrate treated for 10 min nearly doubled compared to the one treated for 5 min. In this work, a fluorescence biosensor was developed to detect breast cancer exosomes using nanoporous gold substrates (NPG). Assay and model exosomes of specific breast cancer cells showed that exosomes exhibit diagnostic surface protein markers, reflecting the protein profile of their parent cells. Furthermore, the specific binding between the exosome surface antibodies and the targets identified the CD63 biomarkers on the exosome, suggesting these markers’ diagnostic potential. This study can accelerate exosome research in determining tumor-related exosomes and develop novel cancer diagnostic methods.
format Online
Article
Text
id pubmed-9460504
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-94605042022-09-10 Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry Al Mannai, Amera Al-Ansari, Tareq Saoud, Khaled M. Sensors (Basel) Article Tumor-derived exosomes may provide biomarkers for cancer treatment. Using sputtering technology, an affinity-based device to capture exosomes was developed using nanoporous substrate (NPG)-coated silicon microscopy. Immunology-based techniques detect and purify exosomes using gold coating with a specific antigen. Inverted fluorescent microscopy was used to detect target exosomes quantitatively utilizing fluorescent nanospheres as the label. We quantified the expression of CD63 surface protein markers on exosomes from conditioned culture media of breast cancer cells. The exosomes that targeted specific proteins with controls were statistically analyzed and compared to those that targeted non-specific proteins. Results from SEM showed that the exosomes were circular, between 30 and 150 nanometers in size. The porous gold substrates captured more exosomes than the nonporous substrates. Nitric acid treatments at different times resulted in a variety of pore sizes. Despite the increase in the size of the pores, the number of exosomes found in the porous gold substrate treated for 10 min nearly doubled compared to the one treated for 5 min. In this work, a fluorescence biosensor was developed to detect breast cancer exosomes using nanoporous gold substrates (NPG). Assay and model exosomes of specific breast cancer cells showed that exosomes exhibit diagnostic surface protein markers, reflecting the protein profile of their parent cells. Furthermore, the specific binding between the exosome surface antibodies and the targets identified the CD63 biomarkers on the exosome, suggesting these markers’ diagnostic potential. This study can accelerate exosome research in determining tumor-related exosomes and develop novel cancer diagnostic methods. MDPI 2022-08-24 /pmc/articles/PMC9460504/ /pubmed/36080806 http://dx.doi.org/10.3390/s22176347 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Al Mannai, Amera
Al-Ansari, Tareq
Saoud, Khaled M.
Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry
title Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry
title_full Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry
title_fullStr Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry
title_full_unstemmed Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry
title_short Quantification of Serum Exosome Biomarkers Using 3D Nanoporous Gold and Spectrophotometry
title_sort quantification of serum exosome biomarkers using 3d nanoporous gold and spectrophotometry
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9460504/
https://www.ncbi.nlm.nih.gov/pubmed/36080806
http://dx.doi.org/10.3390/s22176347
work_keys_str_mv AT almannaiamera quantificationofserumexosomebiomarkersusing3dnanoporousgoldandspectrophotometry
AT alansaritareq quantificationofserumexosomebiomarkersusing3dnanoporousgoldandspectrophotometry
AT saoudkhaledm quantificationofserumexosomebiomarkersusing3dnanoporousgoldandspectrophotometry