Measurement of protein expression using reagents with barcoded oligonucleotide sequences
Some embodiments disclosed herein provide a plurality of compositions each comprising a protein binding reagent conjugated with an oligonucleotide. The oligonucleotide comprises a unique identifier for the protein binding reagent it is conjugated with, and the protein binding reagent is capable of specifically binding to a protein target. Further disclosed are methods and kits for quantitative analysis of a plurality of protein targets in a sample and for simultaneous quantitative analysis of protein and nucleic acid targets in a sample. Also disclosed herein are systems and methods for preparing a labeled biomolecule reagent, including a labeled biomolecule agent comprising a protein binding reagent conjugated with an oligonucleotide.
1. A plurality of compositions each comprising a cellular component binding reagent associated with one or more oligonucleotides, wherein one of the at least one or more oligonucleotides comprises a target region and a unique identifier for the cellular component binding reagent that it is associated therewith, and the cellular component binding reagent is capable of specifically binding to a cellular component target, and the unique identifier is selected from a diverse set of unique identifiers comprising at least 100 different unique identifiers, and wherein the plurality of compositions further comprises one or more second cellular component binding reagents which are not associated with an oligonucleotide.
2. The plurality of compositions of claim 1 , wherein the cellular component target is a carbohydrate, a lipid, a protein, an intracellular protein, an extracellular protein, a cell-surface protein, a cell marker, a B-cell receptor, a T-cell receptor, an antibody, a major histocompatibility complex, a tumor antigen, a receptor, or a combination thereof.
3. The plurality of compositions of claim 1 , wherein the unique identifier sequence comprises a nucleotide sequence of 25-45 nucleotides in length.
4. The plurality of composition of claim 2 , comprising at least 100 compositions each comprising the cellular component binding reagent associated with one or more oligonucleotides.
5. The plurality of composition of claim 4 , wherein at least 10 of the at least 100 compositions comprise different unique identifier sequences.
6. The plurality of composition of claim 5 , wherein at least 100 compositions comprise different unique identifier sequences.
7. The plurality of compositions of claim 1 , wherein the oligonucleotide comprises a barcode sequence, a poly(A) sequence, or a combination thereof.
8. The plurality of compositions of claim 1 , wherein the target region is at the 3′ region of the oligonucleotide.
9. The plurality of composition of claim 1 , wherein the target region comprises a poly(A) sequence.
10. The plurality of compositions of claim 1 , wherein the oligonucleotide is associated to the cellular component binding reagent through a linker.
11. The plurality of compositions of claim 10 , wherein the oligonucleotide comprises the linker.
12. The plurality of compositions of claim 1 , wherein the oligonucleotide is reversibly associated to the cellular component binding reagent.
13. The plurality of compositions of claim 1 , comprising at least 10 compositions each comprising a different cellular component binding reagent.
14. The plurality of compositions of claim 7 , wherein the oligonucleotide associated with the cellular component binding reagent in each composition comprises at least one barcode sequence selected from a set of at least 10 different barcode sequences.
15. The plurality of compositions of claim 1 , wherein the cellular component binding reagent and the one or more second cellular component binding reagents are the same.
16. The plurality of compositions of claim 1 , wherein the cellular component binding reagent is capable of specifically binding to a plurality of cellular component targets.
17. The plurality of compositions of claim 16 , wherein the plurality of cellular component targets comprises a carbohydrate, a lipid, a protein, an intracellular protein, an extracellular protein, a cell-surface protein, a cell marker, a B-cell receptor, a T-cell receptor, an antibody, a major histocompatibility complex, a tumor antigen, a receptor, or any combination thereof.
18. The plurality of compositions of claim 16 , wherein the plurality of cellular component targets comprises 10 to 400 different cellular component targets.
19. A kit for simultaneous quantitative analysis of a plurality of cellular component targets and a plurality of nucleic acid target molecules in a sample, comprising
a plurality of compositions each comprising a cellular component binding reagent associated with one or more oligonucleotides, wherein one of the at least one or more oligonucleotides comprises a target region and a unique identifier for the cellular component binding reagent that it is associated therewith, and the cellular component binding reagent is capable of specifically binding to a cellular component target, and the unique identifier is selected from a diverse set of unique identifiers comprising at least 100 different unique identifiers, and wherein the plurality of compositions further comprises one or more second cellular component binding reagents which are not associated with an oligonucleotide; and
a plurality of oligonucleotide probes, wherein each of the plurality of oligonucleotide probes comprises a target binding region and a barcode sequence, wherein the barcode sequence is from a diverse set of unique barcode sequences.
20. The kit of claim 19 , wherein the cellular component target is a carbohydrate, a lipid, a protein, an intracellular protein, an extracellular protein, a cell-surface protein, a cell marker, a B-cell receptor, a T-cell receptor, an antibody, a major histocompatibility complex, a tumor antigen, a receptor, or a combination thereof.
21. The kit of claim 19 , comprising at least 10 compositions each comprising a different cellular component binding reagent.
22. The kit of claim 19 , comprising at least 100 compositions each comprising the cellular component binding reagent associated with one or more oligonucleotides.
23. The kit of claim 22 , wherein at least 100 compositions comprise different unique identifier sequences.
24. The kit of claim 19 , wherein the oligonucleotide comprises a barcode sequence, a poly(A) sequence, or a combination thereof.
25. The kit of claim 19 , wherein the target region comprises a poly(A) sequence.
26. The plurality of compositions of claim 1 , wherein the cellular component binding reagent comprises an antibody, a tetramer, an aptamer, a protein scaffold, or a combination thereof.
27. The kit of claim 19 , wherein the cellular component binding reagent comprises an antibody, a tetramer, an aptamer, a protein scaffold, or a combination thereof.