US Patent No. 10,921,317

METHODS OF ASSAYING PROTEINS


Patent No. 10,921,317
Issue Date February 16, 2021
Title Methods Of Assaying Proteins
Inventorship Parag Mallick, San Mateo, CA (US)
Assignee Nautilus Biotechnology, Inc., Seattle, WA (US)

Claim of US Patent No. 10,921,317

1. A composition of a protein assay array, the composition comprising:a substrate having a plurality of n protein molecules from a biological sample conjugated to the substrate such that each individual protein of the plurality of n protein molecules is spatially separated from each other protein of the plurality of n protein molecules, and wherein each protein of the plurality of n protein molecules is conjugated at an individual, optically resolvable address, thereby allowing for interrogation of individual protein molecules of the plurality of n protein molecules;
in a first configuration, a first plurality of affinity reagent pools within a liquid medium in communication with the substrate, wherein the liquid medium is in communication with the plurality of n protein molecules conjugated to the substrate, wherein a portion of the affinity reagents within the first plurality of affinity reagent pools is bound or is attached to one or more of the n protein molecules, and wherein at least one affinity reagent within the first plurality of affinity reagent pools recognizes two or more distinct epitopes within the n protein molecules; and
in a second configuration, a second plurality of affinity reagent pools within a liquid medium in communication with the substrate, wherein the liquid medium is in communication with the plurality of n protein molecules conjugated to the substrate, wherein a portion of the affinity reagents within the second plurality of affinity reagent pools is bound or is attached to one or more of the n protein molecules;
wherein the binding of the affinity reagent pools to the plurality of n protein molecules is distinct between the first and second plurality of affinity reagent pools, and wherein the affinity reagents in the affinity reagent pools comprise a known degree of binding nonspecificity to two or more epitopes and are configured to bind to a plurality of epitopes of at least one protein molecule of the plurality of n protein molecules, and wherein the affinity reagents in the affinity reagent pools bind to less than all of the available plurality of epitopes in the plurality of n protein molecules.