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Duplex Microfluidic SERS Detection of Pathogen Antigens with Nanoyeast Single-Chain Variable Fragments
[Image: see text] Quantitative and accurate detection of multiple biomarkers would allow for the rapid diagnosis and treatment of diseases induced by pathogens. Monoclonal antibodies are standard affinity reagents applied for biomarkers detection; however, their production is expensive and labor-int...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American
Chemical
Society
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4188274/ https://www.ncbi.nlm.nih.gov/pubmed/25192256 http://dx.doi.org/10.1021/ac5027012 |
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author | Wang, Yuling Rauf, Sakandar Grewal, Yadveer S. Spadafora, Lauren J. Shiddiky, Muhammad J. A. Cangelosi, Gerard A. Schlücker, Sebastian Trau, Matt |
author_facet | Wang, Yuling Rauf, Sakandar Grewal, Yadveer S. Spadafora, Lauren J. Shiddiky, Muhammad J. A. Cangelosi, Gerard A. Schlücker, Sebastian Trau, Matt |
author_sort | Wang, Yuling |
collection | PubMed |
description | [Image: see text] Quantitative and accurate detection of multiple biomarkers would allow for the rapid diagnosis and treatment of diseases induced by pathogens. Monoclonal antibodies are standard affinity reagents applied for biomarkers detection; however, their production is expensive and labor-intensive. Herein, we report on newly developed nanoyeast single-chain variable fragments (NYscFv) as an attractive alternative to monoclonal antibodies, which offers the unique advantage of a cost-effective production, stability in solution, and target-specificity. By combination of surface-enhanced Raman scattering (SERS) microspectroscopy using glass-coated, highly purified SERS nanoparticle clusters as labels, with a microfluidic device comprising multiple channels, a robust platform for the sensitive duplex detection of pathogen antigens has been developed. Highly sensitive detection for individual Entamoeba histolytica antigen EHI_115350 (limit of detection = 1 pg/mL, corresponding to 58.8 fM) and EHI_182030 (10 pg/mL, corresponding 453 fM) with high specificity has been achieved, employing the newly developed corresponding NYscFv as probe in combination with SERS microspectroscopy at a single laser excitation wavelength. Our first report on SERS-based immunoassays using the novel NYscFv affinity reagent demonstrates the flexibility of NYscFv fragments as viable alternatives to monoclonal antibodies in a range of bioassay platforms and paves the way for further applications. |
format | Online Article Text |
id | pubmed-4188274 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American
Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-41882742015-09-05 Duplex Microfluidic SERS Detection of Pathogen Antigens with Nanoyeast Single-Chain Variable Fragments Wang, Yuling Rauf, Sakandar Grewal, Yadveer S. Spadafora, Lauren J. Shiddiky, Muhammad J. A. Cangelosi, Gerard A. Schlücker, Sebastian Trau, Matt Anal Chem [Image: see text] Quantitative and accurate detection of multiple biomarkers would allow for the rapid diagnosis and treatment of diseases induced by pathogens. Monoclonal antibodies are standard affinity reagents applied for biomarkers detection; however, their production is expensive and labor-intensive. Herein, we report on newly developed nanoyeast single-chain variable fragments (NYscFv) as an attractive alternative to monoclonal antibodies, which offers the unique advantage of a cost-effective production, stability in solution, and target-specificity. By combination of surface-enhanced Raman scattering (SERS) microspectroscopy using glass-coated, highly purified SERS nanoparticle clusters as labels, with a microfluidic device comprising multiple channels, a robust platform for the sensitive duplex detection of pathogen antigens has been developed. Highly sensitive detection for individual Entamoeba histolytica antigen EHI_115350 (limit of detection = 1 pg/mL, corresponding to 58.8 fM) and EHI_182030 (10 pg/mL, corresponding 453 fM) with high specificity has been achieved, employing the newly developed corresponding NYscFv as probe in combination with SERS microspectroscopy at a single laser excitation wavelength. Our first report on SERS-based immunoassays using the novel NYscFv affinity reagent demonstrates the flexibility of NYscFv fragments as viable alternatives to monoclonal antibodies in a range of bioassay platforms and paves the way for further applications. American Chemical Society 2014-09-05 2014-10-07 /pmc/articles/PMC4188274/ /pubmed/25192256 http://dx.doi.org/10.1021/ac5027012 Text en Copyright © 2014 American Chemical Society Terms of Use (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) |
spellingShingle | Wang, Yuling Rauf, Sakandar Grewal, Yadveer S. Spadafora, Lauren J. Shiddiky, Muhammad J. A. Cangelosi, Gerard A. Schlücker, Sebastian Trau, Matt Duplex Microfluidic SERS Detection of Pathogen Antigens with Nanoyeast Single-Chain Variable Fragments |
title | Duplex Microfluidic SERS Detection of Pathogen Antigens
with Nanoyeast Single-Chain Variable Fragments |
title_full | Duplex Microfluidic SERS Detection of Pathogen Antigens
with Nanoyeast Single-Chain Variable Fragments |
title_fullStr | Duplex Microfluidic SERS Detection of Pathogen Antigens
with Nanoyeast Single-Chain Variable Fragments |
title_full_unstemmed | Duplex Microfluidic SERS Detection of Pathogen Antigens
with Nanoyeast Single-Chain Variable Fragments |
title_short | Duplex Microfluidic SERS Detection of Pathogen Antigens
with Nanoyeast Single-Chain Variable Fragments |
title_sort | duplex microfluidic sers detection of pathogen antigens
with nanoyeast single-chain variable fragments |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4188274/ https://www.ncbi.nlm.nih.gov/pubmed/25192256 http://dx.doi.org/10.1021/ac5027012 |
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