Detection of Hepatitis C core antibody by dual-affinity yeast chimera and smartphone-based electrochemical sensing
Yeast cell lines were genetically engineered to display Hepatitis C virus (HCV) core antigen linked to gold binding peptide (GBP) as a dual-affinity biobrick chimera. These multifunctional yeast cells adhere to the gold sensor surface while simultaneously acting as a “renewable” capture reagent for...
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Veröffentlicht in: | Biosensors & bioelectronics 2016-12, Vol.86, p.690-696 |
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Sprache: | eng |
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Zusammenfassung: | Yeast cell lines were genetically engineered to display Hepatitis C virus (HCV) core antigen linked to gold binding peptide (GBP) as a dual-affinity biobrick chimera. These multifunctional yeast cells adhere to the gold sensor surface while simultaneously acting as a “renewable” capture reagent for anti-HCV core antibody. This streamlined functionalization and detection strategy removes the need for traditional purification and immobilization techniques. With this biobrick construct, both optical and electrochemical immunoassays were developed. The optical immunoassays demonstrated detection of anti-HCV core antibody down to 12.3pM concentrations while the electrochemical assay demonstrated higher binding constants and dynamic range. The electrochemical format and a custom, low-cost smartphone-based potentiostat ($20 USD) yielded comparable results to assays performed on a state-of-the-art electrochemical workstation. We propose this combination of synthetic biology and scalable, point-of-care sensing has potential to provide low-cost, cutting edge diagnostic capability for many pathogens in a variety of settings.
•Prospect of a low-cost POC HCV diagnostic test for global healthcare needs.•Yeast dual-affinity reagents based on Aga1-Aga2 surface display expression system.•HCV core protein co-expressed with GBP as a fusion protein to facilitate one-step purification and immobilization.•A $20 US smartphone-based potentiostat for measuring electrochemical assays. |
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ISSN: | 0956-5663 1873-4235 |
DOI: | 10.1016/j.bios.2016.07.023 |