Reagents to Chemically Tag Specific Coronavirus Spike Proteins

Reagents to Chemically Tag Specific Coronavirus Spike Proteins
Firebird Biomolecular Sciences, LLC.Steven A. BennerBharat Gawande
AbstractWhile vaccines and antivirals may be long term solutions to the current coronavirus pandemic, it is now
widely appreciated that the pandemic can be managed before these emerge by rapid, point-of-sampling andquot;public
space entry testsandquot; (PSETs), given appropriate FDA regulatory relief. PSETs will be even more important if a
good vaccine never emerges, a possibility given experience with other coronaviruses.In the 2003 SARS crisis, we delivered a andquot;best in classandquot; PCR kit for SARS that targeted coronaviral RNA. This
was possible due to our first generation nucleic acid innovations, which were also used in respiratory pathogen
panels and cystic fibrosis tests, inter alia. Luminex acquired EraGen in 2011 for $34 million. Since then, the PI
generated 2nd and 3rd generation innovations that supported tests for MERS, arboviruses, norovirus and, this
year, CoV-19 itself. These include a PSET that gives 10 minute results using proprietary displaceable probe loop
amplification. 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 coronavirus
itself. 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. Each
acylation will generate ~300 redox active moieties on an electrode surface per virion. The test is conceptually
advanced 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 andgt;100x sensitivity over Abbott BinaxNOWTM, the assay should detect virus in any contagious individual at
entrances 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 inspike proteins and peptide loops of CoV-19, SARS, and MERS.
Aim 2. Metric the AEGISZymes that emerge, measuring their binding affinities, acylation rates, and specificityamong the 3 homologous surface loops. The metrics are:M1. Acylation with kcat ≈ 1 sec-1. This should be readily achieved based on pmolar affinities that are nowroutine with AEGIS-LIVE; similar rate constants are now routine in analogous catalysts.M2. Specificity with kcat/Kdiss ratios andgt;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 incorporate
AEGISZymes onto its kiosk-style instrument. The kiosk has been proven with standard aptamers to give
electrochemical readouts in less than 1 minute. Firebirdandapos;s AEGISZymes will immediately improve the sensitivity
of 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. BennerBharat Gawande
NarrativeWhile 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 andquot;public space entry testsandquot; (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.