US2022017567A1PendingUtilityA1

Methods of generating nanoarrays and microarrays

Assignee: NAUTILUS BIOTECHNOLOGY INCPriority: Apr 4, 2018Filed: Oct 7, 2021Published: Jan 20, 2022
Est. expiryApr 4, 2038(~11.7 yrs left)· nominal 20-yr term from priority
G01N 33/54353G01N 33/543C12Q 1/6837C07K 1/047
76
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Claims

Abstract

The methods described herein provide a means of producing an array of spatially separated proteins. The method relies on covalently attaching each protein of the plurality of proteins to a structured nucleic acid particle (SNAP), and attaching the SNAPs to a solid support.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A method of producing an array of different proteins, comprising:
 (a) attaching a plurality of structured nucleic acid particles to a plurality of different proteins, such that each of the plurality of structured nucleic acid particles is attached to only one protein of the plurality of different proteins; and   (b) attaching the plurality of structured nucleic acid particles to an array of attachment sites on a solid support, such that each of the array of attachment sites is attached, via a structured nucleic acid particle, to only one protein of the plurality of different proteins, thereby producing the array of different proteins.   
     
     
         2 . The method of  claim 1 , wherein the plurality of structured nucleic acid particles comprises nucleic acid origami. 
     
     
         3 . The method of  claim 2 , wherein the nucleic acid origami comprises a long nucleic acid strand that is hybridized to a plurality of short nucleic acid strands. 
     
     
         4 . The method of  claim 3 , wherein the long nucleic acid strand is hybridized to at least 10 short nucleic acid strands. 
     
     
         5 . The method of  claim 3 , wherein the long nucleic acid strand comprises at least 10 engineered folds. 
     
     
         6 . The method of  claim 3 , wherein the long nucleic acid strand comprises at least 100 nucleotides. 
     
     
         7 . The method of  claim 1 , wherein the structured nucleic acid particle has a diameter between 50 nanometers and 100 micrometers. 
     
     
         8 . The method of  claim 12 , wherein the structured nucleic acid particle has a diameter of at least 75 nanometers. 
     
     
         9 . The method of  claim 1 , wherein the proteins in the array of different proteins are separated by less than 1 micrometers. 
     
     
         10 . The method of  claim 1 , further comprising detecting proteins in the array of different proteins. 
     
     
         11 . The method of  claim 1 , wherein the plurality of structured nucleic acid particles is attached to the array of attachment sites through electrostatic interactions. 
     
     
         12 . The method of  claim 1 , wherein the plurality of structured nucleic acid particles is covalently attached to the array of attachment sites. 
     
     
         13 . The method of  claim 1 , wherein the plurality of structured nucleic acid particles occludes binding of more than one protein to individual attachment sites of the array of attachment sites. 
     
     
         14 . The method of  claim 1 , wherein (a) comprises covalently attaching the plurality of structured nucleic acid particles to the plurality of different proteins, such that each of the plurality of structured nucleic acid particles is covalently attached to only one protein of the plurality of different proteins. 
     
     
         15 . The method of  claim 1 , wherein the array of different proteins comprises 1000×1000 of the plurality of structured nucleic acid particles. 
     
     
         16 . The method of  claim 1 , wherein the plurality of structured nucleic acid particles comprises self-hybridized regions. 
     
     
         17 . The method of  claim 1 , wherein the plurality of structured nucleic acid particles comprises circular plasmids comprising a length of at least 5 kilobases.

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