Electronic methods for the detection of analytes
The present invention is directed to the detection of target analytes using electronic techniques, particularly AC techniques.
1. A method of detecting a protein target analyte in a sample comprising:
a) applying a first input signal to an assay complex comprising:
i) a redox active complex comprising:
1) a redox active molecule;
2) a binding ligand that will bind a protein target analyte; and
ii) said protein target analyte,
wherein said binding ligand is covalently attached to a first measuring electrode via a spacer, and wherein said first measuring electrode comprises:
a passivation agent monolayer comprising at least a covalently attached first passivation species and a covalently attached second passivation species comprising said binding ligand; and
b) detecting a change in the faradaic impedance of the system as a result of the association of the redox active molecule with said protein target analyte, if present.
2. The method according to claim 1 , wherein said input signal comprises an AC component.
3. The method according to claim 1 , wherein said input signal further comprises a DC component.
4. The method according to claim 1 , wherein said redox active molecule is covalently attached to said electrode.
5. The method according to claim 1 , wherein said binding ligand is covalently attached to said electrode.
6. The method according to claim 1 , wherein said redox active molecule is a transition metal complex.
7. The method according to claim 6 , wherein said transition metal complex is ferrocene.
8. The method according to claim 1 , wherein said redox active molecule is covalently attached to said binding ligand.
9. The method according to claim 1 , wherein said detecting is by receiving an output signal characteristic of the presence of said analyte.
10. The method according to claim 9 , wherein said output signal comprises a current.
11. The method according to claim 1 , wherein said conductive oligomer is selected from the group consisting of:
wherein
Y is an aromatic group;
n is an integer from 1 to 50;
g is either 1 or zero;
e is an integer from zero to 10; and
m is zero or 1;
wherein when g is 1, B-D is a conjugated bond; and
wherein when g is zero, e is 1 and D is preferably carbonyl, or a heteroatom moiety, wherein the
heteroatom is selected from oxygen, sulfur, nitrogen, silicon or phosphorus; and
wherein
n is an integer from 1 to 50;
m is 0 or 1;
C is carbon;
J is carbonyl or a heteroatom moiety, wherein the heteroatom is selected from the group consisting of oxygen, nitrogen, silicon, phosphorus, sulfur; and
G is a bond selected from alkane, alkene or acetylene, wherein if m=0, at least one G is not alkane.
12. The method according to claim 1 , wherein said binding ligand comprises a protein.
13. The method according to claim 12 , wherein said protein is a peptide.
14. The method according to claim 1 , wherein said binding ligand comprises a substrate of an enzyme.
15. The method according to claim 1 , wherein said passivation agent monolayer comprises insulators.
16. The method according to claim 1 , wherein said passivation agent monolayer comprises conductive oligomers.
17. The method according to claim 1 , wherein said passivation agent monolayer comprises insulators and conductive oligomers.
18. The method according to claim 15 , wherein said insulator comprises a C l to C12 alkyl group.
19. The method according to claim 1 , wherein binding ligand is covalently attached to said redox active molecule to form a redox active complex.
20. The method according to claim 19 , wherein an atom of said binding ligand serves as a coordination atom to the transition metal to form a redox active complex.
21. The method according to claim 19 , wherein said covalent attachment utilizes a branched linker.
22. The method according to claim 1 , wherein said redox active molecule and said binding ligand are covalently attached to said first measuring electrode separately.