Use of aptamers in proteomics
The present invention is a method for measuring the amount of at least one molecule in a biological sample, the method comprising a) combining the sample, or a derivative thereof, with one or more aptamers and allowing one or more molecules in the sample to bind to the aptamer(s); b) separating bound from unbound molecules; and c) quantifying the molecule(s) bound to the or each aptamer, wherein quantification of the bound molecule(s) is carried out by sequencing at least part of the or each aptamer. Uses of and products derived from the method are also contemplated.
1. A method for detecting or quantifying target molecules within a plurality of molecules in a biological sample, the method comprising:
a) combining the biological sample with a plurality of sets of aptamer clones, wherein each aptamer in each set of aptamer clones is attached to a sequence tag that is unique to that set of aptamer clones;
b) allowing aptamers within the plurality of sets of aptamer clones to bind to target molecules if the target molecules are present within the plurality of molecules in the biological sample;
c) separating aptamers that bound target molecules in step b) from aptamers that did not bind target molecules in step b), and removing the aptamers that did not bind target molecules;
d) sequencing using next generation sequencing techniques the unique sequence tags of each of the aptamers that were not removed in step c) to generate sequencing data; and
e) detecting or quantifying target molecules within the plurality of molecules in the biological sample using the sequencing data generated from the next generation sequencing performed in step d).
2. The method of claim 1 , wherein the identity of the target molecules is known.
3. The method of claim 1 , wherein the identity of the target molecules of each set of aptamer clones is known.
4. The method of claim 1 , wherein the target molecules within the plurality of molecules comprise proteins.
5. The method of claim 1 , wherein the target molecules within the plurality of molecules are identified by proteomics-based analysis.
6. The method of claim 5 , wherein the proteomics-based analysis is high performance liquid chromatography (HPLC) or mass spectrometry (MS).
7. The method of claim 1 , wherein the sequences of the aptamers within the plurality of sets of aptamer clones are known.
8. The method of claim 1 , wherein the sequence tags comprise nucleic acids.
9. The method of claim 8 , wherein the sequence tags are attached to the aptamers using ligation.
10. The method of claim 8 , wherein the sequences of the sequence tags for each of the plurality of sets of aptamer clones are known.
11. The method of claim 8 , wherein the sequencing in step d) comprises sequencing part or all of each sequence tag and part or all of the attached aptamer.
12. The method of claim 1 , further comprising removing part or all of the unique sequence tags from the aptamers prior to the sequencing in step d).
13. The method of claim 1 , wherein the next generation sequencing in step d) comprises a single molecule array or a clonal array.
14. The method of claim 1 , wherein the method further comprises disassociating the aptamers from the target molecules prior to the sequencing in step d).
15. The method of claim 1 , wherein the method further comprises amplifying the sequence tags prior to the sequencing in step d).
16. The method of claim 1 , wherein the plurality of sets of aptamer clones comprises different sets of aptamer clones that bind to the same target molecule.
17. The method of claim 1 , wherein plurality of sets of aptamer clones comprises different sets of aptamer clones that bind to different target molecules.
18. The method of claim 1 , wherein the aptamers within the plurality of aptamers comprise nucleic acids.
19. The method of claim 1 , wherein the plurality of target molecules comprises more than 20 different target molecules.
20. The method of claim 1 , wherein the plurality of target molecules comprises more than 50 different target molecules.
21. The method of claim 1 , wherein the plurality of target molecules comprises more than 100, 500, or 1000 different target molecules.
22. The method of claim 1 , wherein the biological sample comprises urine, saliva, sputum, semen, milk, mucus, sweat, feces, blood, cerebrospinal fluid, interstitial fluid, optic fluid (vitreous), synovial fluid, a tissue biopsy, a resection, or any isolated or enriched portion thereof.
23. The method of claim 1 , wherein the biological sample is obtained from a subject suffering from a condition, disorder, disease or infection.
24. The method of claim 1 , further comprising, prior to the sequencing in step d), adding an adapter sequence to the aptamers that were not removed in step c).
25. The method of claim 1 , further comprising, prior to the sequencing in step d), adding an adapter sequence to the sequence tags of the aptamers that were not removed in step c).
26. The method of claim 24 , wherein part or all of the adapter sequence is sequenced in step d).
27. The method of claim 25 , wherein part or all of the adapter sequence is sequenced in step d).
28. The method of claim 1 , further comprising, prior to the sequencing in step d):
(i) adding one or more adapter sequences to the aptamers that were not removed in step c), wherein the one or more adapter sequences comprise an attachment sequence for attachment of the aptamers to a surface; and
(ii) attaching the aptamers comprising adapter sequences from step i) to a surface.
29. The method of claim 1 , further comprising, prior to the sequencing in step d):
(i) adding one or more adapter sequences to the sequence tags of the aptamers that were not removed in step c), wherein the one or more adapter sequences comprise an attachment sequence for attachment of the sequence tags to a surface; and
(ii) attaching the sequence tags comprising adapter sequences from step i) to a surface.
30. The method of claim 1 , wherein step b) comprises incubating the plurality of sets of aptamer clones with the plurality of molecules in the biological sample at room temperature.