Targeted molecular bar codes and methods for using the same
View Patent ↗Targeted molecular bar codes and methods for using the same are provided. The subject targeted molecular bar codes include a molecular bar code and a member of a specific binding pair, where the specific binding pair member is generally bonded to the bar code through a linking group. The subject molecular bar code may be read during translocation through a single nano-meter scale pore. The subject targeted molecular bar codes find use in a variety of different applications involving analyte detection, such as screening and diagnostic applications.
1. A method for detecting the presence of an analyte in a sample, said method comprising:
(a) contacting said sample with at least one targeted molecular bar code under conditions sufficient for a specific binding pair to bind to said analyte in said sample, wherein said molecular bar code comprises:
(i) a charged polymer capable of generating a reproducible signal upon passage through a nanopore; and
(ii) a member of a specific binding pair, wherein said specific binding pair member is joined directly or through a linking group to said molecular bar code;
(b) separating unbound targeted molecular bar code from analyte-bound targeted molecular bar code;
(c) treating said analyte bound targeted molecular bar code in a manner sufficient to release said molecular bar code from said analyte-bound targeted molecular bar code and produce free molecular bar code;
(e) detecting the presence of said free molecular bar code by translocating said free molecular bar code through a nanopore; and
(f) relating the presence of said free molecular bar code to the presence of said analyte in said sample.
2. The method according to claim 1 , wherein said detecting step further comprises observing a current blockade effect of said translocation on said nanopore.
3. The method according to claim 1 , wherein said method comprises contacting a plurality of different targeted molecular bar codes with said sample.
4. The method according to claim 3 , wherein said sample size does not exceed the size of a biological cell.
5. The method according to claim 1 , wherein said charged polymer is negatively charged.
6. The method according to claim 5 , wherein said negatively charged polymer is made up of monomeric units that comprise a moiety selected from the group consisting of a phosphate group or a phosphorothioate group.
7. The method according to claim 5 , wherein said polymer is a block copolymer of a plurality of blocks, wherein said plurality of blocks are selected from two or more different blocks.
8. The method according to claim 7 , wherein said block copolymer comprises three different blocks.
9. The method according to claim 5 , wherein said molecular bar code comprises a linking group.
10. The method according to claim 9 , wherein said linking group is a photocleavable linking group.
11. A method for detecting the presence of an analyte in a sample, said method comprising:
(a) contacting said sample with at least one targeted molecular bar code under conditions sufficient for a specific binding pair to bind to said analyte in said sample, wherein said molecular bar code comprises:
(i) a negatively charged block copolymer of from one to twenty blocks, wherein said blocks are selected from two or more different blocks, wherein each block consists of monomeric units comprising a phosphate group; and
(ii) a member of a specific binding pair, wherein said member of a specific binding pair is joined to said negatively charged block copolymer through a linking group;
(b) separating unbound targeted molecular bar code from analyte-bound targeted molecular bar code;
(c) treating said analyte bound targeted molecular bar code in a manner sufficient to release said molecular bar code from said analyte-bound targeted molecular bar code and produce free molecular bar code;
(e) detecting the presence of said free molecular bar code; and
(f) relating the presence of said free molecular bar code to the presence of said analyte in said sample.
12. The method according to claim 11 , wherein said detecting step comprises translocating said free molecular bar code through a nanopore, thereby producing a signal.
13. The method according to claim 12 , wherein said detecting step further comprises observing a current blockade effect of said translocation on said nanopore.
14. The method according to claim 11 , wherein said method comprises contacting a plurality of different targeted molecular bar codes with said sample.
15. The method according to claim 14 , wherein said sample size does not exceed the size of a biological cell.
16. The method according to claim 11 , wherein said blocks are selected from two to four different blocks.
17. The method according to claim 11 , wherein each block is a homopolymer of monomeric units selected from the group consisting of phosphates and sugar phosphates.
18. The method according to claim 11 , wherein said sugar phosphates are selected from the group consisting of ribose phosphates and deoxyribose phosphates.
19. The method according to claim 18 , wherein said sugar phosphates may optionally comprise a heterocyclic nitrogenous base.
20. The method according to claim 19 , wherein said heterocyclic nitrogenous base is a purine or a pyrimidine.
21. The method according to claim 11 , wherein the length of each block of said block copolymer ranges from 15 to 25 nm.
22. The method according to claim 21 , wherein said linker is a photocleavable linker.
23. A method for detecting the presence of an analyte in a sample, said method comprising:
(a) contacting said sample with at least one targeted molecular bar code under conditions sufficient for a specific binding pair to bind to said analyte in said sample, wherein said molecular bar code comprises:
(i) a negatively charged block copolymer of from two to twenty blocks, wherein said blocks are selected from a group of three different blocks, wherein each block is a homopolymer of monomeric units selected from the group consisting of polyphosphates, oligonucleotides, oligodeoxyribosephosphates, and polyethylene glycol-phosphodiesters; and
(ii) a member of a specific binding pair, wherein said member of a specific binding pair is joined to said negatively charged block copolymer through a linking group;
(b) separating unbound targeted molecular bar code from analyte-bound targeted molecular bar code;
(c) treating said analyte bound targeted molecular bar code in a manner sufficient to release said molecular bar code from said analyte-bound targeted molecular bar code and produce free molecular bar code;
(e) detecting the presence of said free molecular bar code; and
(f) relating the presence of said free molecular bar code to the presence of said analyte in said sample.
24. The method according to claim 23 , wherein said detecting step comprises translocating said free molecular bar code through a nanopore, thereby producing a signal.
25. The method according to claim 24 , wherein said detecting step further comprises observing a current blockade effect of said translocation on said nanopore.
26. The method according to claim 23 , wherein said method comprises contacting a plurality of different targeted molecular bar codes with said sample.
27. The method according to claim 26 , wherein said sample size does not exceed the size of a biological cell.
28. The method according to claim 23 , wherein said three different blocks are: polyethylene glycol-phosphodiesters; oligodeoxyribosephosphates; and oligonucleotides modified to prevent Watson-Crick base pairing.