There is an urgent need to rapidly detect SARS-CoV-2 (CoV-2) virus in clinical and nonclinical settings, including point of care sites, workplace, and home, at sensitivity and specificity comparable or superior to RT-PCR detection of CoV-2 RNA. Much of the person-to-person CoV-2 transmission occurs before infected individuals develop symptoms. This significant pre-symptomatic/asymptomatic reservoir of CoV-2 transmission mandates efficient identification of infected individuals and their contacts at population-wide screening scale to prevent outbreaks of Covid-19 disease while allowing societies to open and economies to recover. This level of surveillance will also be needed to fully evaluate the effectiveness of countermeasures, including vaccines and therapies. We will develop a new class of diagnostic product reagents, antibody polymers, to create products that can be produced and used at population screening scale for rapid CoV-2 antigen detection at significantly improved sensitivity that approaches the sensitivity of :gold-standard" RT-PCR detection of CoV-2 genomic RNA.Our antibody polymers will be produced by engineering a novel class of small, stable, single polypeptide anti-CoV-2 antibodies termed variable lymphocyte receptors (VLRs) as polymers to achieve essentially irreversible binding to CoV-2 virus. VLR polymers are compatible with all diagnostic immunoassay forms, including lateral flow assay (LFA) "dipsticks", which is likely to be preferred in non-laboratory settings, and with established signaling modalities, e.g., colloidal gold and horseradish peroxidase (HRP). VLRs, the antigen receptors of jawless vertebrates (lamprey and hagfish), are composed of highly diverse leucine-rich repeat domains and arethe only known antigen-specific immune receptors that are not immunoglobulins (Igs). The binding site of VLRsis contained within a small single polypeptide and comprised by amino acid residues in the rigid beta-sheets that form the concave surface of the VLR structure. The over 500 million year evolutionary separation of jawed and jawless vertebrates and distinctive antigen-binding site structure of VLR antibodies have proved a source of novel specificities distinct from conventional Ig antibodies. The outer envelope of the CoV-2 virus is composed of a multivalent array of spike (S) protein that is an ideal target(s) for multivalent, essentially irreversible binding, by appropriately multivalent binding agents. We will genetically link in tandem genes encoding VLRs with binding specificity for CoV-2 S protein to create such multivalent VLR polymer binding agents that couple binding of CoV-2 antigens to an amplifiable, visually observable result, e.g., color change. The high ratio of CoV-2 S protein 50- 100 S protein trimers per virus particle, to the single copy CoV-2 RNA genome, and the amplification available via catalysis, e.g., HRP, provides significantly improved CoV-2 antigen detection sensitivity that approaches sensitivity achieved with RT-PCR detection of single copy CoV-2 RNA.
Project narrative: We will develop a new class of diagnostic reagents, antibody polymers, to create low cost, rapid, easy to use SARS-CoV-2 (CoV-2) antigen detection products with sensitivity that approaches that of "gold-standard" RT-PCR and that can be produced and used at population screening scale. Our antibody polymers will be produced by engineering a novel class of small, stable, single polypeptide anti-CoV-2 antibodies termed variable lymphocyte receptors (VLRs) as polymers to achieve essentially irreversible binding to CoV-2 virus. Our VLR polymers are compatible with all diagnostic immunoassay forms, including lateral flow assay (LFA) "dipsticks", which is likely to be preferred in non-laboratory setting, e.g., point-of-care, workplace and home. Amino Acids ; aminoacid ; Antibodies ; Antigens ; immunogen ; Binding Sites ; Combining Site ; Reactive Site ; Biomedical Research ; Catalysis ; Gold Colloid ; Colloidal Gold ; Color ; Disease ; Disorder ; Engineering ; Epidemiology ; epidemiologic ; epidemiological ; Genes ; Genome ; Gold ; Hagfish ; Horseradish Peroxidase ; Human ; Modern Man ; Immune Sera ; Antisera ; immune serum ; Immunoassay ; Infection ; Jaw ; Lampreys ; Lamprey Eels ; Petromyzontidae ; Libraries ; Lymphocyte ; Lymphatic cell ; Lymphocytic ; lymph cell ; Persons ; Polymers ; Proteins ; Reagent ; Antigen Receptors ; Fc Receptor ; antibody receptor ; Immunologic Receptors ; Immunological Receptors ; immune receptor ; Research ; RNA ; Non-Polyadenylated RNA ; RNA Gene Products ; Ribonucleic Acid ; viral RNA ; virus RNA ; Sensitivity and Specificity ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; Societies ; Specificity ; Vaccines ; Vertebrates ; Vertebrate Animals ; vertebrata ; Virion ; Virus Particle ; Virus ; Yeasts ; Caring ; Workplace ; Job Location ; Job Place ; Job Setting ; Job Site ; Work Location ; Work Place ; Work-Site ; Worksite ; work setting ; base ; community planning ; improved ; Site ; Surface ; Solid ; Clinical ; Phase ; Link ; Individual ; disease transmission ; communicable disease transmission ; infectious disease transmission ; Diagnostic ; Immunes ; Immune ; Source ; Receptor Protein ; receptor ; RT-PCR ; RTPCR ; reverse transcriptase PCR ; Reverse Transcriptase Polymerase Chain Reaction ; Beta Sheet ; β-Sheet ; β-pleated sheet ; beta pleated sheet ; Cellulose Nitrate ; Nitrocellulose ; Pyroxylin ; Structure ; novel ; Modality ; disease control ; disorder control ; Manufacturer ; Manufacturer Name ; antigen bound ; antigen binding ; Molecular Interaction ; Binding ; preventing ; prevent ; polypeptide ; Receptor Gene ; Symptoms ; Detection ; Diagnostic Reagent ; Leucine-Rich Repeat ; Right-Handed Beta-Alpha Superhelix ; Small Business Innovation Research Grant ; SBIR ; Small Business Innovation Research ; transmission process ; Transmission ; point of care ; Behavioral ; cost ; viral detection ; virus detection ; rapid detection ; Population ; Coupled ; phase 2 study ; phase II study ; screening ; genomic RNA ; Immunize ; COVID-19 ; COVID19 ; CV-19 ; CV19 ; corona virus disease 2019 ; coronavirus disease 2019 ; 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 ; effectiveness evaluation ; assess effectiveness ; determine effectiveness ; effectiveness assessment ; evaluate effectiveness ; 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 ; lateral flow assay ; lateral flow test ; antigen detection ; antigen based detection ; detect antigen ; detection sensitivity ; COVID-19 outbreak ; COVID19 outbreak ; SARS-CoV-2 outbreak ; SARS-CoV2 outbreak ; Severe acute respiratory syndrome coronavirus 2 outbreak ; coronavirus disease 2019 outbreak ; Home ;
