SBIR-STTR Award

Direct to Phase II

Direct bioelectronic detection of SARS-CoV-2 from saliva using single-molecule field-effect transistor array Nucleic acid tests have become the gold-standard for diagnostic testing for COVID-19, usually performedin specialized laboratories. Most are based on reverse-transcription quantitative polymerase chain reaction(qRT-PCR). The time required for specimen transport and processing results in a turnaround time that is typi-cally several days. The few rapid (<1 hour) point-of-care (POC) tests are more expensive, still require samplepreparation and specialized reagents, and do not have the throughput needed for population surveillance. Di-rect testing for the virus, which also reduces requirements for multiple reagents, is a necessary step to improv-ing diagnostic testing. While four such antigen tests have been approved for detection of SARS-CoV-2 basedon immunoassays to the N protein, sensitivity is limited and no quantitation of viral load is possible. We will address this gap by using DiagnostikosTM, an in-development rapid POC platform for direct, real-time, multiplexed, quantitative bioelectronic detection of biomolecules that employs an all-electronic detectiondevice that functions at the single-molecule level. These single-molecule field-effect transistors (smFETs) arearrayed on a complementary metal-oxide-semiconductor (CMOS) integrated circuit chip. Chips will interfacewith an envisioned USB-stick-form-factor reader device. Robust single-domain antibodies, known as nanobod-ies and immobilized on these devices, are used for sensitive detection of viral particles and viral debris. Theuse of multiple nanobodies for a single protein and nanobodies for different proteins in a single assay allowsfor significant improvements in specificity. Nanobodies will be specific for one or more of the four major struc-tural proteins in SARS-CoV-2; the nucleocapsid (N) protein engulfing the viral RNA, the spike (S) protein, themembrane (M) protein and the envelope (E) protein. No sample preparation or specialized reagents are re-quired for detection, and the device will be designed to operate with saliva, which has very recently beenshown to be a reliable medium for detecting SARS-CoV-2. Individual sensor chips can be manufactured at acost of $35. With the addition of other nanobodies, these large dense arrays can also allow detection of manypathogens in a single test. In this Direct-To-Phase-2 SBIR program we will pursue several key innovations that are required to makesuch a platform possible, including isolation of nanobodies for key structure proteins of SARS-CoV-2 (SpecificAim 1), development of the smFET platform for antigen detection (Specific Aim 2), development of largeCMOS arrays of these smFET devices (Specific Aim 3), and verification of detection in increasingly complexsamples up to and including clinical samples (Specific Aim 4). This project is a partnership between universityresearchers who developed the smFET technology and a venture-based start-up venture, Quicksilver Biosci-ences, spun out to commercialize smFET technology and develop smFET/CMOS arrays for molecular diag-nostic applications.

Public Health Relevance Statement:
Public Health Narrative The proposal seeks to provide a new bioelectronic approach for antigen testing for infectious diseases. Low- cost and pervasive antigen testing would be an essential new tool for the control of the COVID-19 pandemic.

Project Terms:
Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Biological Sciences ; Biologic Sciences ; Bioscience ; Life Sciences ; Communicable Diseases ; Infectious Disease Pathway ; Infectious Diseases ; Infectious Disorder ; Diagnosis ; Differential Diagnosis ; Electrolytes ; env Gene Products ; Envelope Protein ; env Antigens ; env Polyproteins ; env Protein ; Gold ; Immobilization ; orthopedic freezing ; Immunoassay ; Infection ; Influenza ; Grippe ; Kinetics ; Laboratories ; Llama ; Membrane Proteins ; Membrane Protein Gene ; Membrane-Associated Proteins ; Surface Proteins ; New York ; Optics ; optical ; Polymerase Chain Reaction ; Population Surveillance ; Public Health Surveillance ; Proteins ; Public Health ; Reagent ; Research Personnel ; Investigators ; Researchers ; Reverse Transcription ; viral RNA ; virus RNA ; Saliva ; Semiconductors ; Sensitivity and Specificity ; Specificity ; Technology ; Testing ; Time ; Transistors ; Universities ; Vaccines ; Viral Proteins ; Viral Gene Products ; Viral Gene Proteins ; virus protein ; Virus ; Diagnostic tests ; base ; sensor ; improved ; Nucleic Acid Amplification Tests ; Nucleic Acid Testing ; Site ; Clinical ; Phase ; Training ; Individual ; Binding Proteins ; Ligand Binding Protein ; Ligand Binding Protein Gene ; Protein Binding ; bound protein ; infectious organism ; Infectious Agent ; tool ; Research Specimen ; Specimen ; programs ; Viral Burden ; Viral Load ; Viral Load result ; Hour ; Complex ; Viral ; Nucleocapsid Proteins ; metal oxide ; particle ; Devices ; Sampling ; molecular array ; single molecule ; ImProv ; Buckytubes ; Carbon nano tubes ; Carbon Nanotubes ; nano tubes ; Nanotubes ; Address ; Defect ; Symptoms ; protein structures ; proteins structure ; protein structure ; Detection ; Quantitative RTPCR ; qRTPCR ; Quantitative Reverse Transcriptase PCR ; Reader ; Recombinants ; Small Business Innovation Research Grant ; SBIR ; Small Business Innovation Research ; transmission process ; Transmission ; Preparation ; Process ; Development ; developmental ; point of care ; cost ; multiplex detection ; viral detection ; virus detection ; design ; designing ; pathogen ; innovation ; innovate ; innovative ; biobank ; biorepository ; nanobodies ; nanobody ; sdAb ; single domain antibodies ; temporal measurement ; temporal resolution ; time measurement ; diagnostic assay ; molecular diagnostics ; Structural Protein ; experimental study ; experiment ; experimental research ; 2019-nCoV ; 2019 novel corona virus ; 2019 novel coronavirus ; COVID-19 virus ; COVID19 virus ; CoV-2 ; CoV2 ; SARS corona virus 2 ; SARS-CoV-2 ; SARS-CoV2 ; SARS-associated corona virus 2 ; SARS-associated coronavirus 2 ; SARS-coronavirus-2 ; SARS-related corona virus 2 ; SARS-related coronavirus 2 ; SARSCoV2 ; Severe Acute Respiratory Distress Syndrome CoV 2 ; Severe Acute Respiratory Distress Syndrome Corona Virus 2 ; Severe Acute Respiratory Distress Syndrome Coronavirus 2 ; Severe Acute Respiratory Syndrome CoV 2 ; Severe Acute Respiratory Syndrome-associated coronavirus 2 ; Severe Acute Respiratory Syndrome-related coronavirus 2 ; Severe acute respiratory syndrome associated corona virus 2 ; Severe acute respiratory syndrome corona virus 2 ; Severe acute respiratory syndrome coronavirus 2 ; Severe acute respiratory syndrome related corona virus 2 ; Wuhan coronavirus ; coronavirus disease 2019 virus ; hCoV19 ; nCoV2 ; COVID-19 pandemic ; COVID crisis ; COVID epidemic ; COVID pandemic ; COVID-19 crisis ; COVID-19 epidemic ; COVID-19 global health crisis ; COVID-19 global pandemic ; COVID-19 health crisis ; COVID-19 public health crisis ; COVID19 crisis ; COVID19 epidemic ; COVID19 global health crisis ; COVID19 global pandemic ; COVID19 health crisis ; COVID19 pandemic ; COVID19 public health crisis ; SARS-CoV-2 epidemic ; SARS-CoV-2 global health crisis ; SARS-CoV-2 global pandemic ; SARS-CoV-2 pandemic ; SARS-CoV2 epidemic ; SARS-CoV2 pandemic ; SARS-coronavirus-2 epidemic ; SARS-coronavirus-2 pandemic ; Severe Acute Respiratory Syndrome CoV 2 epidemic ; Severe Acute Respiratory Syndrome CoV 2 pandemic ; Severe acute respiratory syndrome coronavirus 2 epidemic ; Severe acute respiratory syndrome coronavirus 2 pandemic ; corona virus disease 2019 epidemic ; corona virus disease 2019 pandemic ; coronavirus disease 2019 crisis ; coronavirus disease 2019 epidemic ; coronavirus disease 2019 global health crisis ; coronavirus disease 2019 global pandemic ; coronavirus disease 2019 health crisis ; coronavirus disease 2019 pandemic ; coronavirus disease 2019 public health crisis ; coronavirus disease crisis ; coronavirus disease epidemic ; coronavirus disease pandemic ; severe acute respiratory syndrome coronavirus 2 global health crisis ; severe acute respiratory syndrome coronavirus 2 global pandemic ; COVID-19 diagnostic ; COVID19 diagnostic ; SARS-CoV-2 diagnostic ; coronavirus disease 2019 diagnostic ; severe acute respiratory syndrome coronavirus 2 diagnostic ; COVID test ; COVID tests ; coronavirus disease test ; coronavirus disease tests ; coronavirus test ; coronavirus tests ; COVID-19 testing ; COVID19 testing ; SARS-CoV-2 testing ; coronavirus disease 2019 testing ; severe acute respiratory syndrome coronavirus 2 testing ; COVID-19 detection ; COVID19 detection ; SARS-CoV-2 detection ; coronavirus disease 2019 detection ; detect COVID-19 ; detect COVID19 ; detect SARS-CoV-2 ; detect severe acute respiratory syndrome coronavirus 2 ; severe acute respiratory syndrome coronavirus 2 detection ; saliva sample ; salivary sample ; antigen test ; antigen based test ; antibody detection ; antibody based detection ; detect antibodies ; antigen detection ; antigen based detection ; detect antigen ; SARS-CoV-2 transmission ; COVID-19 transmission ; COVID-19 virus transmission ; coronavirus disease 2019 transmission ; coronavirus disease 2019 virus transmission ; severe acute respiratory syndrome coronavirus 2 transmission ; transmitted COVID-19 ; transmitted SARS-CoV-2 ; transmitted coronavirus disease 2019 ; transmitted severe acute respiratory syndrome coronavirus 2 ; point of care testing ;