Macromolecule analysis employing nucleic acid encoding

1. A method for analyzing peptides of a single cell from a sample comprising a population of cells, the method comprising the following steps:

(a) partitioning cells of the population of cells into a plurality of compartments, wherein:

i) a single cell comprising peptides and at least one bead from a plurality of beads are comprised within a single compartment; and

ii) each bead of the plurality of beads comprises a plurality of nucleic acid recording tags attached thereto, wherein each nucleic acid recording tag of the plurality of nucleic acid recording tags comprises a barcode specific for each compartment and a functional moiety configured to covalently link the nucleic acid recording tag to a peptide of the peptides of the single cell, thereby immobilizing the peptide on the bead via the nucleic acid recording tag attached thereto;

(b) lysing cells in the compartments and allowing immobilization of the peptides released from the single cell on the at least one bead comprised within the single compartment, thereby generating at least one bead comprising immobilized barcoded peptides of the single cell, wherein each of the immobilized barcoded peptides is associated with a nucleic acid recording tag of the plurality of nucleic acid recording tags;

(c) releasing the beads comprising the immobilized barcoded peptides from the plurality of compartments and pooling the released beads; and

(d) analyzing the immobilized barcoded peptides on the released beads, wherein 10,000 or more immobilized barcoded peptides are analyzed simultaneously, and the analyzing comprises: (i) contacting the released beads with a plurality of binding agents capable of binding to the immobilized barcoded peptides; and (ii) following binding of a binding agent from the plurality of binding agents to an immobilized barcoded peptide, obtaining information regarding the binding agent, thereby analyzing the immobilized barcoded peptides of the single cell.

2. The method of claim 1 , wherein prior to partitioning the cells into the compartments, the cells are encapsulated in droplets containing polymer-forming subunits, and prior to immobilization of the peptides released from the single cell on the at least one bead, the peptides are digested with a protease, and peptides produced by the protease digestion are immobilized on the at least one bead.

3. The method of claim 1 , wherein prior to immobilization of the peptides released from the single cell on the at least one bead, the peptides are digested with a protease, and peptides produced by the protease digestion are immobilized on the at least one bead.

4. The method of claim 1 , wherein prior to partitioning the cells into the compartments or prior to immobilization of peptides on the beads, the peptides are contacted with a labeling reagent that is configured to provide the peptides with a reactive click chemistry coupling moiety capable of reacting with the functional moiety of the nucleic acid recording tags attached to the beads, and wherein the reactive click chemistry coupling moiety reacts with the nucleic acid recording tags via a click chemistry reaction.

5. The method of claim 4 , wherein prior to contacting with the labeling reagent, the peptides are denatured.

6. The method of claim 4 , wherein prior to immobilization of the peptides released from the single cell on the at least one bead, the peptides are digested with a protease, and peptides produced by the protease digestion are immobilized on the at least one bead.

7. The method of claim 1 , wherein 100 or more different immobilized barcoded peptides are analyzed simultaneously in step (d).

8. The method of claim 7 , wherein analyzing the immobilized barcoded peptides on the released beads further comprises determining at least partial sequences of the immobilized barcoded peptides.

9. The method of claim 7 , wherein analyzing the immobilized barcoded peptides on the released beads further comprises the following steps:

(e) contacting the released beads with a plurality of binding agents capable of binding to the immobilized peptides, wherein each binding agent comprises a coding tag with identifying information regarding the binding agent;

(f) after binding of a binding agent to an immobilized peptide, transferring information of the coding tag with identifying information regarding the binding agent to the nucleic acid recording tag associated with the immobilized peptide to generate an extended nucleic acid recording tag associated with the immobilized peptide, wherein the transferring comprises primer extension or ligation; and

(g) analyzing a plurality of the extended nucleic acid recording tags generated after transfer of information from the plurality of binding agents, thereby analyzing peptides of the single cell.

10. The method of claim 9 , wherein the plurality of nucleic acid extended recording tags is analyzed in parallel by performing a nucleic acid sequencing method.

11. The method of claim 9 , wherein on each bead of the plurality of beads, the nucleic acid recording tags are spaced apart from each other on a surface or within a volume of the bead by at least 50 nm.

12. The method of claim 9 , wherein at least one binding agent from the plurality of binding agents is capable of binding to a N-terminal amino acid (NTAA) of an immobilized barcoded peptide, or is capable of binding to a N-terminal amino acid (NTAA) of an immobilized barcoded peptide modified with a small chemical moiety.

13. The method of claim 9 , wherein steps (e) and (f) are repeated at least once before step (g).

14. The method of claim 1 , wherein the functional moiety configured to immobilize the peptides comprises a peptide substrate motif of a protein ligase, and the method further comprises adding the protein ligase to the plurality of compartments, thereby coupling the peptides to the beads.

15. The method of claim 1 , wherein the functional moiety configured to immobilize the peptides is a reactive click chemistry coupling moiety, an aldehyde, or a peptide motif for a protein ligase.

16. The method of claim 1 , wherein 500 or more different immobilized barcoded peptides are analyzed simultaneously in step (d).