Detection of conductive polymer-labeled analytes
View Patent ↗The disclosure relates to the detection of analytes (e.g., biological pathogens such as bacteria or viruses) using a conductive polymer label. The disclosed detection system utilizing the conductive polymer label generally involves the formation of an analyte conjugate between the target analyte and a conductive polymer moiety conjugated to the target analyte. The conductive polymer portion of the analyte conjugate is electrically activated to form an electrically activated analyte conjugate having an increased electrical conductivity relative to the analyte conjugate as originally formed. The electrically activated analyte conjugate can then be detected by any suitable means, such as by conductimetric or electrochemical detection.
1. A method for detecting the presence of a target analyte, the method comprising:
(a) providing an analyte conjugate comprising: (i) the target analyte and (ii) a conductive polymer moiety conjugated to the target analyte via a binding pair member bound to the conductive polymer moiety and capable of binding to the target analyte;
(b) electrically activating the analyte conjugate by acid-doping the analyte conjugate, thereby forming an electrically activated analyte conjugate having an increased electrical conductivity relative to the analyte conjugate; and
(c) detecting the electrically activated analyte conjugate.
2. The method of claim 1 , wherein the analyte conjugate comprises:
(i) a particulate composition comprising a conductive polymer as the conductive polymer moiety bound to a magnetic nanoparticle,
(ii) the binding pair member bound to the conductive polymer of the particulate composition, the binding pair member being capable of specifically binding to the target analyte, and
(iii) the target analyte bound to the binding pair member.
3. The method of claim 1 , wherein the analyte conjugate comprises:
(i) a magnetic nanoparticle capture composition comprising: (A) a magnetic nanoparticle, and (B) a first binding pair member bound to the magnetic nanoparticle, the first binding pair member being capable of specifically binding to the target analyte,
(ii) a conductive polymer nanoparticle label composition comprising: (A) a conductive polymer nanoparticle as the conductive polymer moiety, and (B) a second binding pair member bound to the conductive polymer nanoparticle as the binding pair member bound to the conductive polymer moiety, the second binding pair member being capable of specifically binding to the target analyte, and
(iii) the target analyte bound to both the first binding pair member and the second binding pair member.
4. The method of claim 1 , wherein the analyte conjugate is provided in a liquid aqueous medium having a pH value of 5 or more, and the analyte conjugate is electrically activated in the liquid aqueous medium.
5. The method of claim 1 , further comprising, before detecting the electrically activated analyte conjugate, immobilizing the analyte conjugate or the electrically activated analyte conjugate on a detection surface of a biosensor via at least one of a specific or non-specific binding pair interaction and a magnetic interaction therebetween.
6. The method of claim 5 , comprising immobilizing the analyte conjugate on the detection surface before electrically activating the analyte conjugate.
7. The method of claim 5 , comprising immobilizing the electrically activated analyte conjugate on the detection surface.
8. The method of claim 5 , wherein:
(i) the detection surface of the biosensor has opposing top and bottom surfaces, where the electrically activated analyte conjugate is immobilized on the top surface;
(ii) the biosensor further comprises a magnetic means for generating a magnetic field positioned adjacent the bottom surface of the detection surface;
(iii) the electrically activated analyte conjugate further comprises a magnetic moiety conjugated to the target analyte or bound to the conductive polymer of the electrically activated analyte conjugate; and
(iv) immobilizing the analyte conjugate or the electrically activated analyte conjugate comprises (A) generating the magnetic field with the magnetic means to magnetically position the analyte conjugate or the electrically activated analyte conjugate closer to the top surface of the detection surface than in the absence of the magnetic field, and (B) maintaining the magnetic field when detecting the electrically activated analyte conjugate.
9. The method of claim 8 , wherein:
(i) the biosensor comprises a biosensor binding pair member capable of specific or non-specific binding to the target analyte, the biosensor binding pair member being immobilized on the detection surface; and
(ii) immobilizing the analyte conjugate or the electrically activated analyte conjugate comprises (A) first immobilizing the analyte conjugate or the electrically activated analyte conjugate on the detection surface through a binding interaction between the binding pair member and the target analyte of the analyte conjugate or the electrically activated analyte conjugate, (B) washing the detection surface in the absence of a magnetic field, (C) generating the magnetic field with the magnetic means to magnetically position the analyte conjugate or the electrically activated analyte conjugate closer to the top surface of the detection surface than in the absence of the magnetic field, and (D) maintaining the magnetic field when detecting the electrically activated analyte conjugate.
10. The method of claim 5 , wherein the biosensor is a screen-printed carbon electrode (SPCE), and the detection surface is a working electrode of the SPCE.
11. The method of claim 1 , further comprising: determining that the target analyte is present in a sample from which the analyte conjugate is formed.
12. The method of claim 1 , wherein the target analyte is selected from the group consisting of a bacterium and a virus.