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 measuring the amount of at least one molecule in a biological sample, the method comprising:
a) combining the biological sample, or a derivative thereof, with a plurality of aptamers and allowing members of the plurality of aptamers to bind to the molecules in the sample;
b) separating molecules that are bound by members of the plurality of aptamers in step a); and
c) sequencing, using next generation polynucleotide sequencing techniques, at least part of each of the members of the plurality of aptamers that bound the molecules separated in step b) to quantify each of the bound aptamers,
thereby measuring the amount of the at least one molecule.
2. The method of claim 1 , wherein the at least one molecule comprises a protein.
3. The method of claim 1 , wherein the identity of the at least one molecule is known.
4. The method of claim 1 , wherein the identity of the at least one molecule is unknown.
5. The method of claim 4 , wherein the method further comprises determining the identity of the at least one molecule.
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 the sequence of each unique member of the plurality of aptamers carries a unique tag.
8. The method of claim 7 , wherein the tag comprises the sequence of each member of the plurality of aptamers.
9. The method of claim 7 , wherein the tag comprises part of the sequence of each member of the plurality of aptamers.
10. The method of claim 1 , wherein the next generation polynucleotide 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 arraying the members of the plurality of aptamers that bound the molecules separated in step b) onto a surface.
12. The method of claim 11 , wherein the method further comprises amplifying the arrayed members of the plurality of aptamers.
13. The method of claim 1 , wherein the plurality of aptamers comprises different member sequences that bind to the 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 comprises DNA, RNA, or both.
16. The method of claim 1 , wherein the members of the plurality of aptamers are between about 30 and about 60 bases long.
17. The method of claim 1 , wherein the members of the plurality of aptamers are about 40 bases long.
18. The method of claim 1 , wherein the biological sample comprises a bodily fluid.
19. The method of claim 18 , wherein the bodily fluid comprises blood or is derived from blood.
20. The method of claim 18 , wherein the bodily fluid comprises serum or plasma.
21. The method of claim 1 , wherein the method further comprises:
d) combining a second biological sample with each of the members of the plurality of aptamers that bound the molecules separated in step b);
e) separating molecules in the second biological sample that are bound by members of the plurality of aptamers in step d);
f) sequencing, using next generation polynucleotide sequencing techniques, at least part of each of the members of the plurality of aptamers that bound the molecules separated in step e), thereby measuring the amount of the at least one molecule in the second biological sample; and
g) comparing the amounts of the at least one molecule obtained in c) with those obtained in f).
22. The method of claim 1 , wherein the method further comprises comparing the quantity of the members of the plurality of aptamers sequenced in step c) against a control or baseline quantity.
23. The method of claim 21 , wherein the second biological sample is obtained from an individual known to be in a diseased state.
24. The method of claim 21 , wherein the second biological sample is obtained from an individual known to be in a healthy state.
25. The method of claim 21 , wherein the second biological sample is obtained from an individual after drug treatment.
26. The method of claim 22 , further comprising diagnosing a disease or predicament based on the comparison.