Rapid antifouling nanocomposite coating enables highly sensitive multiplexed electrochemical detection of myocardial infarction and concussion markers

S, Sharma Timilsina; Durr, Nolan; Yafia, Mohamed; Sallum, Hani; Jolly, Pawan; Ingber, Donald E.

doi:10.1101/2021.06.13.21258856

Cited by 4 publications

(17 citation statements)

References 34 publications

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“…More recently, using rapid coating of BSA/prGOx/GA, we accomplished multiplexed detection for multiple MI and TBI biomarkers in undiluted whole blood and plasma. 3 EC sensor assays were developed for six different biomarkers of different sizes, including B-type natriuretic peptide (BNP; 3.5 kDa) and its N-terminal fragment (NT-proBNP; 8.5 kDa), S100b (12 kDa), cardiac troponin I (cTnI; 34.6 kDa), glial fibrillary acidic protein (GFAP; 50 kDa), and cTnI-TC complex; 77.5 kDa. We reached single-digit pg/ mL sensitivity using only 15 μL of unprocessed plasma and whole blood samples for all of these biomarkers (Figure 4C).…”

Section: Multiplexing Strategiessupporting

confidence: 82%

“…In addition, the multiplexed EC sensor platform was validated by analyzing patient samples and demonstrating excellent correlation with reported clinical values. 2,3 Finally, because of the high sensitivity and selectivity of the nanocomposite coated EC sensors, we were able to develop a high-density multiplexed sensor array to carry out simultaneous detection of multiple clinically relevant MI and TBI biomarkers using unprocessed human plasma samples. Four different specific capture probes (cTnl-TC, S100b, NT-proBNP, and GFAP) were individually functionalized on each sensing electrode (Figure 4D).…”

Section: Multiplexing Strategiesmentioning

confidence: 99%

“…61 However, by combining these nanoparticles with polymers and proteins efficient antifouling characteristics can be achieved. 2,3 To evaluate stability of our platform, we first explored the shelf life of the EC biosensor as bioreceptors bound to the antifouling layer may degrade or detach over time. Electrodes functionalized with BSA/AuNW/GA-coating and anti-IL-6 antibodies were successfully regenerated after storage for up to 1 month in 1% BSA, we could detect 200 pg/mL IL-6 with minimum loss in current density, confirming the long-term stability of antifouling coated EC biosensor (Figure 3B).…”

Section: Stability Of the Nanocomposite Coatingmentioning

confidence: 99%

“…2 This describes an EC sensor platform that enables the detection of multiple sepsis biomarkers simultaneously by incorporating a nanocomposite coating composed of cross-linked bovine serum albumin containing a network of reduced graphene oxide nanoparticles that prevents biofouling while maintaining electroconductivity. 3 This describes an ultrafast (<1 min) method for coating multiplexed electrochemical sensors with an antifouling nanocomposite layer that can be stored at room temperature for months, which provides unprecedented sensitivity and selectivity in whole blood for diagnostic applications using clinically relevant biomarkers.…”

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confidence: 99%

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Enabling Multiplexed Electrochemical Detection of Biomarkers with High Sensitivity in Complex Biological Samples

Timilsina¹,

Jolly²,

Durr³

et al. 2021

Acc. Chem. Res.

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Conspectus The ability to perform multiplexed detection of various biomarkers within complex biological fluids in a robust, rapid, sensitive, and cost-effective manner could transform clinical diagnostics and enable personalized healthcare. Electrochemical (EC) sensor technology has been explored as a way to address this challenge because it does not require optical instrumentation and it is readily compatible with both integrated circuit and microfluidic technologies; yet this approach has had little impact as a viable commercial bioanalytical tool to date. The most critical limitation hindering their clinical application is the fact that EC sensors undergo rapid biofouling when exposed to complex biological samples (e.g., blood, plasma, saliva, urine), leading to the loss of sensitivity and selectivity. Thus, to break through this barrier, we must solve this biofouling problem. In response to this challenge, our group has developed a rapid, robust, and low-cost nanocomposite-based antifouling coating for multiplexed EC sensors that enables unprecedented performance in terms of biomarker signal detection compared to reported literature. The bioinspired antifouling coating that we developed is a nanoporous composite that contains various conductive nanomaterials, including gold nanowires (AuNWs), carbon nanotubes (CNTs), or reduced graphene oxide nanoflakes (rGOx). Each study has progressively evolved this technology to provide increasing performance while simplifying process flow, reducing time, and decreasing cost. For example, after successfully developing a semipermeable nanocomposite coating containing AuNWs cross-linked to bovine serum albumin (BSA) using glutaraldehyde, we replaced the nanomaterials with reduced graphene oxide, reducing the cost by 100-fold while maintaining similar signal transduction and antifouling properties. We, subsequently, developed a localized heat-induced coating method that significantly improved the efficiency of the drop-casting coating process and occurs within the unprecedented time of <1 min (at least 3 orders of magnitude faster than state-of-the-art). Moreover, the resulting coated electrodes can be stored at room temperature for at least 5 months and still maintain full sensitivity and specificity. Importantly, this improved coating showed excellent antifouling activity against various biological fluids, including plasma, serum, whole blood, urine, and saliva. To enable affinity-based sensing of multiple biomarkers simultaneously, we have developed multiplexed EC sensors coated with the improved nanocomposite coating and then employed a sandwich enzyme-linked immunosorbent assay (ELISA) format for signal detection in which the substrate for the enzyme bound to the secondary antibody precipitates locally at the molecular binding site above the electrode surface. Using this improved EC sensor platform, we demonstrated ultrasensitive detection of a wide range of biomarkers from biological fluids, including clinical biomarkers, in both single and multiplex formats (N = 4) wi...

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Section: Multiplexing Strategiessupporting

confidence: 82%

Section: Multiplexing Strategiesmentioning

confidence: 99%

Section: Stability Of the Nanocomposite Coatingmentioning

confidence: 99%

mentioning

confidence: 99%

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Enabling Multiplexed Electrochemical Detection of Biomarkers with High Sensitivity in Complex Biological Samples

Timilsina¹,

Jolly²,

Durr³

et al. 2021

Acc. Chem. Res.

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“…Due to its customizable surface chemistry, these EC sensors enable the detection of different targets such as nucleic acids and proteins. The simple and inexpensive BSA/rGOx/GA-based surface chemistry of the antifouling sensor coating combined with a poly-HRP streptavidin/TMB output allows for high conductivity and low nonspecific binding, leading to ultra-low EC background and improved sensitivity 37,60 .…”

Section: Discussionmentioning

confidence: 99%

Lab-on-a-chip multiplexed electrochemical sensor enables simultaneous detection of SARS-CoV-2 RNA and host antibodies

Puig

Timilsina

Rainbow

et al. 2021

Preprint

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There continues to be a great need for rapid, accurate, and cost-effective point-of-care devices that can diagnose the presence of SARS-CoV-2 virus and development of IgG and IgM antibody responses in early and late stages of COVID-19 disease. Here, we describe a versatile multiplexed electrochemical (EC) sensor platform modified with an antifouling nanocomposite coating that enables single-molecule CRISPR/Cas-based molecular detection of SARS-CoV-2 viral RNA with on-chip signal validation as well as multiplexed serological detection of antibodies against three SARS-CoV-2 viral antigens. The CRISPR-based EC platform achieved 100% accuracy for detection of viral RNA and showed an excellent correlation with RT-qPCR using 30 clinical saliva samples. The serology EC platform obtained 100% sensitivity and 100% specificity for anti-SARS-CoV-2 IgG, as well as 94% specificity and 82% sensitivity for anti-SARS-CoV-2 IgM with 112 clinical plasma samples. These data demonstrate that integration of CRISPR-based RNA detection and serological assays with antifouling nanocomposite-based EC sensors enables performance as good or better than traditional laboratory-based techniques.

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A lab-on-a-chip for the concurrent electrochemical detection of SARS-CoV-2 RNA and anti-SARS-CoV-2 antibodies in saliva and plasma

et al. 2022

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Rapid, accurate and frequent detection of the RNA of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) and of serological host antibodies to the virus would facilitate the determination of the immune status of individuals who have Coronavirus disease 2019 (COVID-19), were previously infected by the virus, or were vaccinated against the disease. Here we describe the development and application of a 3D-printed lab-on-a-chip that concurrently detects, via multiplexed electrochemical outputs and within 2 h, SARS-CoV-2 RNA in saliva as well as anti-SARS-CoV-2 immunoglobulins in saliva spiked with blood plasma. The device automatedly extracts, concentrates and amplifies SARS-CoV-2 RNA from unprocessed saliva, and integrates the Cas12a-based enzymatic detection of SARS-CoV-2 RNA via isothermal nucleic acid amplification with a sandwich-based enzyme-linked immunosorbent assay on electrodes functionalized with the Spike S1, nucleocapsid and receptor-binding-domain antigens of SARS-CoV-2. Inexpensive microfluidic electrochemical sensors for performing multiplexed diagnostics at the point of care may facilitate the widespread monitoring of COVID-19 infection and immunity.

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Rapid antifouling nanocomposite coating enables highly sensitive multiplexed electrochemical detection of myocardial infarction and concussion markers

Cited by 4 publications

References 34 publications

Enabling Multiplexed Electrochemical Detection of Biomarkers with High Sensitivity in Complex Biological Samples

Enabling Multiplexed Electrochemical Detection of Biomarkers with High Sensitivity in Complex Biological Samples

Lab-on-a-chip multiplexed electrochemical sensor enables simultaneous detection of SARS-CoV-2 RNA and host antibodies

A lab-on-a-chip for the concurrent electrochemical detection of SARS-CoV-2 RNA and anti-SARS-CoV-2 antibodies in saliva and plasma

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