Duplex microfluidic SERS detection of pathogen antigens with nanoyeast single-chain variable fragments

Yuling Wang, Sakandar Rauf, Yadveer S. Grewal, Lauren J. Spadafora, Muhammad J.A. Shiddiky, Gerard A. Cangelosi, Sebastian Schlücker, Matt Trau

Research output: Contribution to journalArticlepeer-review

59 Citations (Scopus)
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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 EHI115350 (limit of detection = 1 pg/mL, corresponding to 58.8 fM) and EHI182030 (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.

Original languageEnglish
Pages (from-to)9930-9938
Number of pages9
JournalAnalytical Chemistry
Issue number19
Publication statusPublished - 07 Oct 2014


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