Electrochemical aptamer sensor monolayer incubation with improved stability

1 . A method of fabricating an electrochemical aptamer sensor comprising:

incubating an electrode in the presence of components that are adapted to constitute at least a portion of a sensing monolayer, wherein the sensing monolayer includes at least a plurality of aptamers and at least one weakly bonded constituent;

removing the at least one weakly bonded constituent from the sensing monolayer by applying at least one perturbation mechanism to the sensing monolayer while incubating the electrode, or by incubating the electrode then applying at least one perturbation mechanism to the sensing monolayer then incubating the electrode again;

wherein the plurality of aptamers each comprise at least one redox tag, and applying the perturbation mechanism results in a normalized current measured as a peak signal from the redox tag versus a background current decrease over 24 hours that is an amount selected from the group consisting of less than 3%, less than 5%, less than 10%, less than 20%, and less than 40%.

2 . The method of claim 1 , wherein the perturbation mechanism is selected from the group consisting of an electrical mechanism, a mechanical mechanism, a chemical mechanism, a thermal mechanism, and a combination thereof.

3 . The method of claim 1 , wherein the sensing monolayer further comprises a blocking layer.

4 . The method of claim 3 , wherein the blocking layer comprises a material that is an alkythiolate.

5 . The method of claim 1 , wherein the plurality of aptamers are at least one of greater than 30%, greater than 60%, greater than 90%, or greater than 95% chemically attached to the electrode compared to a total amount of aptamers on the electrode surface via both chemical and physiosorbed attachment.

6 . An electrochemical aptamer sensor comprising:

an electrode having an electrode surface;

a sensing monolayer, the sensing monolayer comprising at least a plurality of molecules forming the sensing monolayer, the sensing monolayer attached to the electrode surface;

wherein the plurality of molecules attached to the electrode surface may be either weakly bonded or strongly bonded to the electrode surface;

and wherein, of the plurality of molecules, at least one of less than 40%, less than 20%, less than 10% or less than 5% of the sensing monolayer are weakly bonded to the electrode surface.

7 . The sensor of claim 6 , wherein at least a portion of the molecules are aptamers.

8 . The sensor of claim 6 , wherein at least a portion of the molecules form a blocking layer.

9 . A method of fabricating an electrochemical aptamer sensor comprising:

incubating an electrode in the presence of components that are adapted to constitute at least a portion of a sensing monolayer, wherein the sensing monolayer includes at least a plurality of aptamers and at least one weakly bonded constituent;

removing the at least one weakly bonded constituent from the sensing monolayer by applying at least one perturbation mechanism to the sensing monolayer while incubating the electrode, or by incubating the electrode then applying at least one perturbation mechanism to the sensing monolayer then incubating the electrode again;

wherein the plurality of aptamers are at least one of greater than 30%, greater than 60%, greater than 90%, or greater than 95% chemically attached to the electrode compared to a total amount of aptamers on the electrode surface via both chemical and physiosorbed attachment.

10 . The method of claim 9 , wherein the perturbation mechanism is selected from the group consisting of an electrical mechanism, a mechanical mechanism, a chemical mechanism, a thermal mechanism, and a combination thereof.

11 . The method of claim 9 , wherein the sensing monolayer further comprises a blocking layer.

12 . The method of claim 11 , wherein the blocking layer comprises a material that is an alkythiolate.