E-DNA scaffold sensors and the reagentless, single-step, measurement of HIV-diagnostic antibodies in human serum

• Published in: Microsystems & nanoengineering Vol. 6; no. 1; p. 13
• Main Authors: Parolo, Claudio, Greenwood, Ava S., Ogden, Nathan E., Kang, Di, Hawes, Chase, Ortega, Gabriel, Arroyo-Currás, Netzahualcóyotl, Plaxco, Kevin W.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 23.03.2020; Springer Nature B.V
• Subjects: 639/638; 639/925/350/59; Deoxyribonucleic acid; Diagnostic systems; DNA; Electron transfer; Engineering; Enzyme-linked immunosorbent assay; Epitopes; HIV; Human immunodeficiency virus; Immunoglobulins; Monoclonal antibodies; Multiplexing; Quantitation; Scaffolds; Sensitivity; Sensors; Steric hindrance
• ISSN: 2055-7434; 2096-1030; 2055-7434
• DOI: 10.1038/s41378-019-0119-5

The multiplexed, point-of-care measurement of specific antibodies could improve the speed with which diseases are diagnosed and their treatment initiated. To this end, we are developing E-DNA scaffold sensors, which consist of a rigid, nucleic acid “scaffold” attached on one end to an electrode and presenting both a redox reporter and an epitope on the other. In the absence of antibody, the reporter efficiently transfers electrons when interrogated electrochemically. Binding-induced steric hindrance limits movement, reducing electron transfer in a manner that is both easily measured and quantitatively related to target concentration. Previously we have used monoclonal antibodies to explore the analytical performance of E-DNA sensors, showing that they support the rapid, single-step, quantitative detection of multiple antibodies in small volume samples. Here, in contrast, we employ authentic human samples to better explore the platform’s clinical potential. Specifically, we developed E-DNA sensors targeting three HIV-specific antibodies and then compared the analytical and clinical performance of these against those of gold standard serological techniques. Doing so we find that, although the multistep amplification of an ELISA leads to a lower detection limits, the clinical sensitivity of ELISAs, E-DNA sensors and lateral-flow dipsticks are indistinguishable across our test set. It thus appears that, by merging the quantitation and multiplexing of ELISAs with the convenience and speed of dipsticks, E-DNA scaffold sensors could significantly improve on current serological practice. Sensors: detecting HIV antibodies A single-step, electrochemical-based device has the ability to detect and measure the amount of HIV antibodies in clinical samples. There is an ever-growing need for tools that can diagnose diseases accurately, quickly and at point-of-care. The group of Kevin Plaxco at University of California Santa Barbara has previously demonstrated an E-DNA scaffold sensor for quantifying antibodies, but it had not been tested on real clinical samples. Now, they demonstrate that it can detect several antibodies for the diagnosis of HIV in clinical samples, and benchmark it against traditional gold standard techniques. The E-DNA sensor achieves 90% sensitivity and 100% specificity for the tested antibodies, values identical to those obtained using standard ELISA and lateral flow immunoassays when challenged with the same sample set.