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 and Trau, Matt (2014) Duplex microfluidic SERS detection of pathogen antigens with nanoyeast single-chain variable fragments. Analytical Chemistry, 86 19: 9930-9938. doi: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
Title Duplex microfluidic SERS detection of pathogen antigens with nanoyeast single-chain variable fragments
Journal name Analytical Chemistry   Check publisher's open access policy
ISSN 0003-2700
1520-6882
Publication date 2014-09-05
Year available 2014
Sub-type Article (original research)
DOI 10.1021/ac5027012
Open Access Status
Volume 86
Issue 19
Start page 9930
End page 9938
Total pages 9
Place of publication Washington, DC, United States
Publisher American Chemical Society
Collection year 2015
Language eng
Formatted abstract
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.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Forthcoming: "Article ASAP".

 
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Created: Fri, 26 Sep 2014, 09:29:54 EST by Jon Swabey on behalf of Aust Institute for Bioengineering & Nanotechnology