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.
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.