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 identifying biomarkers of disease states and/or drug treatments, the method comprising:
(a) combining a biological sample, or a derivative thereof, with a plurality of aptamers and allowing members of the plurality of aptamers to bind to their target molecules in the biological sample;
(b) separating members of the plurality of aptamers that bound target molecules in step a) from members of the plurality of aptamers that did not bind target molecules in step a);
(c) using next generation sequencing techniques to sequence at least part of each of the members of the plurality of aptamers that bound the target molecules in step b) to identify and/or quantify each of the bound aptamers;
(d) determining the identity of the target molecules of the bound aptamers and generating a profile of the target molecules;
(e) comparing the profile of the target molecules with a control profile from a control or baseline sample; and
(f) identifying target molecules whose occurrence or expression levels are altered between the biological sample and the control or baseline sample, thereby identifying biomarkers of disease states and/or drug treatments.
2. The method of claim 1 , wherein the target molecules of the aptamers comprise proteins.
3. The method of claim 1 , wherein the identity of the target molecules of the aptamers is known.
4. The method of claim 1 , wherein the target molecules are identified by proteomics-based analysis.
5. The method of claim 4 , wherein the proteomics-based analysis is high performance liquid chromatography (HPLC) or mass spectrometry (MS).
6. The method of claim 1 , wherein the sequence of the members of the plurality of aptamers is known.
7. The method of claim 1 , wherein each unique aptamer sequence carries a unique tag.
8. The method of claim 7 , wherein the tag is the sequence of each unique aptamer.
9. The method of claim 7 , wherein the tag is part of the sequence of each unique aptamer.
10. The method of claim 1 , wherein the next generation sequencing is carried out on a single molecule array or a clonal single molecule array.
11. The method of claim 1 , wherein the method further comprises disassociating the bound aptamers and molecules separated in step b) prior to step c).
12. The method of claim 1 , wherein the method further comprises amplifying the plurality of aptamers in step b) prior to step c).
13. The method of claim 1 , wherein the plurality of aptamers comprises different member sequences that each bind to a same target molecule.
14. The method of claim 1 , wherein the plurality of aptamers comprises different panels of aptamer sequences that each bind to a different target molecule.
15. The method of claim 1 , wherein the members of the plurality of aptamers comprise DNA, RNA or both.
16. The method of claim 15 , wherein the members of the plurality of aptamers are between about 30 and about 60 bases.
17. The method of claim 1 , wherein the control sample is a biological sample obtained from a healthy individual.
18. The method of claim 1 , wherein the control sample is a biological sample obtained from an individual known to have cancer, or an individual after drug treatment.
19. The method of claim 1 , further comprising the steps of:
(i) adding an adapter sequence to each of the members of the plurality of aptamers that bound the target molecules in step b), wherein the adapter sequence comprises an attachment sequence for attachment of the aptamers to a surface; and
(ii) attaching the members of the plurality of aptamers comprising an adapter sequence from step c) to a surface.
20. The method of claim 1 , wherein the biological sample is blood, tissue biopsy or resection.