While vaccines and antivirals may be long term solutions to the current coronavirus pandemic, it is nowwidely appreciated that the pandemic can be managed before these emerge by rapid, point-of-sampling "publicspace entry tests" (PSETs), given appropriate FDA regulatory relief. PSETs will be even more important if agood vaccine never emerges, a possibility given experience with other coronaviruses. In the 2003 SARS crisis, we delivered a "best in class" PCR kit for SARS that targeted coronaviral RNA. Thiswas possible due to our first generation nucleic acid innovations, which were also used in respiratory pathogenpanels and cystic fibrosis tests, inter alia. Luminex acquired EraGen in 2011 for $34 million. Since then, the PIgenerated 2nd and 3rd generation innovations that supported tests for MERS, arboviruses, norovirus and, thisyear, CoV-19 itself. These include a PSET that gives 10 minute results using proprietary displaceable probe loopamplification. Intrinsic chemistry suggests, however, this is the fastest any RNA-targeted test can be. Here, we will use our 3rd-generation DNA innovations not to target coronaviral RNA, but the coronavirusitself. Firebird will create reagents (AEGISZymes) from an artificially expanded genetic information system(AEGIS) that catalyze covalent acylation of lysines on the surface of ~300 spike proteins per virus. Eachacylation will generate ~300 redox active moieties on an electrode surface per virion. The test is conceptuallyadvanced over antibody tests, as it uses stable covalent bond formation, rather than non-covalent antibody:antigen interactions, and exploits electrochemical detection rather than a lateral flow. Our Phase 2 partner,MightyGate, has shown with standard aptamers detection in less than a minute with such architectures.With >100x sensitivity over Abbott BinaxNOWTM, the assay should detect virus in any contagious individual atentrances to arenas, schools, and other spaces as fast has handbags and are checked. Specifically, we will:Aim 1. Under IRB 2020001, we will use AEGIS-LIVE to evolve AEGISZymes in saliva that acylate lysines in spike proteins and peptide loops of CoV-19, SARS, and MERS.Aim 2. Metric the AEGISZymes that emerge, measuring their binding affinities, acylation rates, and specificity among the 3 homologous surface loops. The metrics are: M1. Acylation with kcat â 1 sec-1. This should be readily achieved based on pmolar affinities that are now routine with AEGIS-LIVE; similar rate constants are now routine in analogous catalysts. M2. Specificity with kcat/Kdiss ratios >103, to ensure that CoV-19 is distinguished from other coronaviruses. If the metrics are met, MightyGate has committed itself to participate in Phase 2 work that will incorporateAEGISZymes onto its kiosk-style instrument. The kiosk has been proven with standard aptamers to giveelectrochemical readouts in less than 1 minute. Firebird's AEGISZymes will immediately improve the sensitivityof that kiosk, and immediate commercial value. However, our approach is general to any virus target. Reagents to Chemically Tag Specific Coronavirus Spike Proteins
Firebird Biomolecular Sciences, LLC.
Steven A. Benner
Bharat Gawande
Narrative
While vaccines and antivirals may be long term solutions to the current coronavirus pandemic, coronaviruses
have been difficult to manage using vaccines, and antiviral drugs are slow and difficult to develop. Fortunately,
it is now widely appreciated that the pandemic can be managed before these emerge, and even if they never
emerge, by rapid, point-of-sampling "public space entry tests" (PSETs). This project will deliver reagents that
tag covalently coronaviral proteins for use in a kiosk-style PSET instrument that our partner, MightyGate, has
shown to be able to generate true/false readouts in less than a minute, the time it takes to check a backpack
or handbag. Once in place, this product will allow the economy to open free of fear of coronavirus infection in
public spaces that are protected by the products of this research. Acylation ; Antibodies ; Antigens ; immunogen ; Antiviral Agents ; Antiviral Drugs ; Antivirals ; anti-viral agents ; anti-viral drugs ; anti-virals ; Arboviruses ; Arboviral ; Arthropod-Borne Viruses ; Architecture ; Engineering / Architecture ; Asia ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Biological Sciences ; Biologic Sciences ; Bioscience ; Life Sciences ; Chemistry ; Clinical Laboratory Information Systems ; Laboratory Information Systems ; Crystallization ; Cystic Fibrosis ; Mucoviscidosis ; Disulfides ; DNA ; Deoxyribonucleic Acid ; Electrodes ; Environment ; Europe ; Evolution ; Fright ; Fear ; Genes ; In Vitro ; India ; Information Systems ; Data Systems ; IT Systems ; Information Technology Systems ; instrumentation ; Laboratories ; Lung diseases ; Pulmonary Diseases ; Pulmonary Disorder ; Respiratory Disease ; Respiratory System Disease ; Respiratory System Disorder ; disease of the lung ; disorder of the lung ; lung disorder ; Lysine ; L-Lysine ; Molecular Conformation ; Molecular Configuration ; Molecular Stereochemistry ; conformation ; conformational state ; Nucleic Acids ; Nucleotides ; Oxidation-Reduction ; Redox ; oxidation reduction reaction ; Peptides ; Protein Engineering ; genetic protein engineering ; protein design ; Proteins ; Publishing ; Reagent ; Research ; RNA ; Non-Polyadenylated RNA ; RNA Gene Products ; Ribonucleic Acid ; Saliva ; Schools ; Science ; Specificity ; Testing ; Time ; Toxin ; Vaccines ; Virion ; Virus Particle ; Virus ; Work ; Generations ; Measures ; catalyst ; base ; improved ; Procedures ; Lateral ; Surface ; Phase ; Medical ; Coronavirus ; Coronaviridae ; corona virus ; Coronavirus Infections ; Coronaviridae Infections ; Ensure ; Chemicals ; Individual ; Binding Proteins ; Ligand Binding Protein ; Ligand Binding Protein Gene ; Protein Binding ; bound protein ; analog ; Coronavirus spike protein ; CoV S protein ; CoV glycoprotein S ; CoV spike glycoprotein ; CoV spike protein ; Coronavirus glycoprotein S ; coronavirus S protein ; coronavirus spike glycoprotein ; Malignant Cell ; cancer cell ; tool ; Hepatic Cancer ; liver cancer ; malignant liver tumor ; Malignant neoplasm of liver ; instrument ; Diagnostic ; Reaction ; amino group ; covalent bond ; disulfide bond ; experience ; nuclease ; Performance ; aptamer ; Structure ; Cell surface ; Regulation ; Sampling ; response ; Norwalk-like Viruses ; Norovirus ; Molecular Interaction ; Binding ; SARS ; SARS coronavirus disease ; SARS-CoV disease ; Severe Acute Respiratory Syndrome CoV disease ; Severe Acute Respiratory Syndrome coronavirus disease ; Severe Acute Respiratory Syndrome ; Amino Acylation ; Aminoacylation ; Affinity ; Detection ; breast tumor cell ; Breast Cancer Cell ; Reproducibility ; Preparation ; Development ; developmental ; pandemic disease ; pandemic ; Image ; imaging ; innovation ; innovate ; innovative ; Institutional Review Boards ; IRB ; IRBs ; genetic information ; symptom treatment ; symptomatic treatment ; treat symptom ; recruit ; novel coronavirus ; CoV emergence ; corona virus emergence ; coronavirus emergence ; emergent CoV ; emergent corona virus ; emergent coronavirus ; emerging CoV ; emerging corona virus ; emerging coronavirus ; nCoV ; new CoV ; new corona virus ; new coronavirus ; novel CoV ; novel corona virus ; 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 ; Middle East Respiratory Syndrome ; MERS ; MERS coronavirus disease ; MERS-CoV disease ; Middle East Respiratory Syndrome CoV disease ; Middle East Respiratory Syndrome coronavirus disease ; Middle Eastern Respiratory Syndrome ; Middle Eastern Respiratory Syndrome CoV disease ; Middle Eastern Respiratory Syndrome coronavirus disease ; respiratory pathogen ; lung pathogen ; pulmonary pathogen ; COVID detection ; coronavirus detection ; coronavirus disease detection ; detect COVID ; detect coronavirus ; detect coronavirus disease ; antibody test ; antibody based test ;
