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US 12,742,168 B2Granted
Systems and methods for biomolecule retention
AssigneeNautilus Subsidiary, Inc.
InventorsTorri Elise Rinker, Christina Inman, Steven Tan, Pengyu Hao, Elvis Ikwa, Pierre Indermuhle, Parag Mallick, Tural Aksel, Stephen Hendricks, Sadie Ingle
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Abstract
Compositions, systems, and methods for the display of analytes such as biomolecules are described. Display of analytes is achieved by coupling of the analytes to displaying molecules that are configured to associate with surfaces or interfaces. Arrays of analytes may be formed from the described systems for utilization in assays and other methods.
Claims28
A method, comprising:
- (a) providing a solid support comprising a plurality of sites, wherein each individual site of the plurality of sites comprises:
- (i) only one particle coupled to the individual site;
- and
- (ii) only one polypeptide of interest attached to the only one particle;
- wherein a first site of the plurality of sites is attached to a first proteoform of a species of polypeptide of interest, and wherein a second site of the plurality of sites is attached to a second proteoform of the species of polypeptide of interest;
- (b) delivering first binding reagents to the solid support, thereby coupling a binding reagent of the first binding reagents to the first proteoform at the first site;
- (c) delivering second binding reagents to the solid support, thereby coupling a binding reagent of the second binding reagents to the second proteoform at the second site;
- and
- (d) detecting at single-analyte resolution a presence of the first binding reagent at the first site and detecting at single-analyte resolution a presence of the second binding reagent at the second site, thereby detecting the first proteoform and the second proteoform, wherein the first proteoform and the second proteoform are different.
6 more elements
A method, comprising:
- (a) providing a solid support comprising a plurality of sites, wherein each individual site of the plurality of sites comprises:
- (i) only one particle coupled to the individual site;
- and
- (ii) only one polypeptide of interest attached to the only one particle;
- (b) delivering first binding reagents to the solid support, wherein the first binding reagents have a binding specificity for a first post-translational modification (PTM), thereby coupling one or more binding reagents of the first binding reagents to one or more polypeptides of interest;
- (c) delivering second binding reagents to the solid support, wherein the second binding reagents have a binding specificity for a second PTM, thereby coupling one or more binding reagents of the second binding reagents to one or more polypeptides of interest;
- and
- (d) for each site of the plurality of sites, detecting at single-analyte resolution a presence or absence of the first binding reagent and the second binding reagent, thereby detecting one or more proteoforms of the polypeptides of interest.
5 more elements
A method, comprising:
- (a) providing a solid support comprising a plurality of wells, wherein each individual well of the plurality of wells comprises:
- (i) only one particle coupled within the individual well;
- and
- (ii) only one polypeptide of interest attached to the only one particle;
- wherein a first well of the plurality of wells contains a first proteoform of a species of polypeptide of interest, and wherein a second well of the plurality of wells contains a second proteoform of the species of polypeptide of interest;
- (b) delivering first binding reagents to the solid support, thereby coupling a binding reagent of the first binding reagents to the first proteoform at the first well;
- (c) delivering second binding reagents to the solid support, thereby coupling a binding reagent of the second binding reagents to the second proteoform at the second well;
- and
- (d) detecting at single-analyte resolution a presence of the first binding reagent at the first well and detecting at single-analyte resolution a presence of the second binding reagent at the second well, thereby detecting the first proteoform and the second proteoform, wherein the first proteoform and the second proteoform are different.
6 more elements
Dependencies are inferred from the claim text.