US 12,320,813 B2
Macromolecule analysis employing nucleic acid encoding
Mark S. Chee, Encinitas, CA (US); Kevin L. Gunderson, Encinitas, CA (US); and Michael Phillip Weiner, San Diego, CA (US)
Assigned to Encodia, Inc., San Diego, CA (US)
Filed by Encodia, Inc., San Diego, CA (US)
Filed on May 9, 2024, as Appl. No. 18/659,334.
Application 18/659,334 is a division of application No. 17/197,796, filed on Mar. 10, 2021, granted, now 12,019,077.
Application 17/197,796 is a continuation of application No. 16/098,436, granted, now 12,123,878, previously published as PCT/US2017/030702, filed on May 2, 2017.
Claims priority of provisional application 62/376,886, filed on Aug. 18, 2016.
Claims priority of provisional application 62/339,071, filed on May 19, 2016.
Claims priority of provisional application 62/330,841, filed on May 2, 2016.
Prior Publication US 2024/0295563 A1, Sep. 5, 2024
Int. Cl. C40B 40/10 (2006.01); C40B 70/00 (2006.01); G01N 33/68 (2006.01)
CPC G01N 33/6824 (2013.01) [C40B 40/10 (2013.01); C40B 70/00 (2013.01); G01N 33/6803 (2013.01); G01N 33/6842 (2013.01); G01N 33/6845 (2013.01); G01N 2458/00 (2013.01); G01N 2458/10 (2013.01)] 9 Claims
 
5. A method for analyzing peptides of a single cell from a sample comprising a population of cells, the method comprising:
(a) partitioning cells of the population of cells into a plurality of compartments such that a single cell comprising peptides is comprised within a single compartment, wherein the compartment comprises polymer-forming subunits;
(b) lysing cells in the compartments, polymerizing the polymer-forming subunits to form single cell gel beads and allowing immobilization of the peptides from the single cell inside a single cell gel bead within the compartment, thereby generating at least one single cell gel bead comprising immobilized peptides of the single cell;
(c) releasing the single cell gel beads comprising the immobilized peptides from the plurality of compartments and pooling the released beads;
(d) barcoding the immobilized peptides on the released single cell gel beads via combinatorial indexing through a series of split-and-pool steps by adding a plurality of recording tags, wherein each recording tag of the plurality of recording tags comprises a barcode and a functional moiety configured to covalently link the recording tag to an immobilized peptide of the immobilized peptides, thereby generating at least one single cell gel bead comprising immobilized barcoded peptides of the single cell; and
(e) analyzing the immobilized barcoded peptides on the released single cell gel beads, thereby analyzing peptides of the single cell.