Macromolecule analysis employing nucleic acid encoding
A method for analyzing macromolecules, including peptides, polypeptides, and proteins, employing nucleic acid encoding is disclosed.
1. A composition comprising:
(a) one or more solid supports, wherein each solid support comprises at least 100,000 peptide molecules covalently attached to the solid support, wherein
(i) each of the at least 100,000 peptide molecules is associated with a nucleic acid tag;
(ii) the at least 100,000 peptide molecules comprise molecules of at least 100 different peptides; and
(iii) adjacent peptide molecules on the solid support are spaced apart from each other on a surface or within a volume of the solid support at an average distance of about 50 nm or greater; and
(b) a plurality of binding agents each bound to one of the peptide molecules, wherein the plurality of binding agents comprises at least 3 different binding agents, wherein each of the at least 3 different binding agents is (i) bound to a N-terminal amino acid (NTAA) residue or C-terminal amino acid (CTAA) residue of a peptide molecule of the peptide molecules, and (ii) selective for a different peptide component, wherein the different peptide component is a different N-terminal amino acid (NTAA) residue or a different C-terminal amino acid (CTAA) residue of the peptide molecules.
2. The composition of claim 1 , wherein each peptide molecule is covalently attached to the nucleic acid tag associated therewith.
3. The composition of claim 1 , wherein the peptide molecules are randomly attached to the solid support.
4. The composition of claim 1 , wherein each peptide molecule is covalently attached to the solid support through a nucleic acid molecule.
5. The composition of claim 4 , wherein the nucleic acid molecule is covalently attached to the C-terminal amino acid residue of the peptide molecule.
6. The composition of claim 4 , wherein the nucleic acid molecule comprises the nucleic acid tag associated with the peptide molecule.
7. The composition of claim 1 , wherein the at least 100,000 peptide molecules comprise at least 1000 different peptide molecules each covalently attached to the support and associated with a nucleic acid tag.
8. The composition of claim 1 , wherein each peptide molecule comprises an N-terminal or C-terminal amino acid residue modified by a chemical agent.
9. The composition of claim 1 , wherein each peptide molecule comprises an N-terminal amino acid (NTAA) residue modified by a chemical agent.
10. The composition of claim 1 , wherein the nucleic acid tag associated with each peptide molecule comprises a barcode or a unique molecular identifier (UMI).
11. The composition of claim 10 , wherein the barcode comprises an analyte barcode, a compartment barcode, a partition barcode, a sample barcode, a fraction barcode, or any combination thereof.
12. The composition of claim 10 , wherein the nucleic acid tags associated with the peptide molecules share a common sequence.
13. The composition of claim 10 , wherein the common sequence comprises a universal primer sequence and a common spacer sequence flanking the barcode or UMI.
14. The composition of claim 1 , wherein the one or more supports are one or more beads or one or more flow cells.
15. The composition of claim 14 , comprising at least 500,000 peptide molecules covalently attached to the same bead.
16. The composition of claim 1 , wherein each binding agent of the plurality of binding agents comprises a coding tag that comprises an encoder sequence comprising identifying information regarding the binding agent.
17. The composition of claim 1 , wherein the plurality of binding agents comprises an aptamer, an antibody, an anticalin, a modified or variant aminopeptidase, a modified or variant amino acyl tRNA synthetase, a modified or variant anticalin, or a modified or variant ATP-dependent Clp protease adaptor protein (ClpS).
18. The composition of claim 10 , wherein each of the at least 100 different peptides is associated with a different nucleic acid tag.
19. The composition of claim 10 , wherein the at least 100,000 peptide molecules are obtained by fragmenting proteins from a biological sample.
20. The composition of claim 19 , wherein at least some of the at least 100,000 peptide molecules comprise post-translational modifications.
21. The composition of claim 20 , wherein the post-translational modifications comprise glycosylation.
22. The composition of claim 1 , wherein the different peptide component is a different NTAA residue of the peptide molecules, and each of the at least 3 different binding agents is selective for a different non-modified NTAA residue of the peptide molecules.
23. The composition of claim 1 , wherein the different peptide component is a different CTAA of the peptide molecules, and each of the at least 3 different binding agents is selective for a different non-modified CTAA residue of the peptide molecules.
24. The composition of claim 1 , wherein each of the plurality of binding agents comprises a polypeptide or protein.
25. The composition of claim 1 , wherein each of the plurality of binding agents is non-covalently bound to one of the peptide molecules.
26. The composition of claim 16 , wherein the coding tag of the binding agent that is bound to a peptide molecule of the at least 100,000 peptide molecules hybridizes to a complementary sequence on the nucleic acid tag associated with the peptide molecule.