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An electrochemical method and apparatus of identifying the presence of a target

An electrode and single-electron technology, applied in biochemical equipment and methods, electrochemical variables of materials, measuring devices, etc., can solve problems such as rapid emergence of resistance

Inactive Publication Date: 2012-05-16
UNIV OF WESTERN ONTARIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Unfortunately, resistance to these drugs emerges rapidly during infection

Method used

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  • An electrochemical method and apparatus of identifying the presence of a target
  • An electrochemical method and apparatus of identifying the presence of a target
  • An electrochemical method and apparatus of identifying the presence of a target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 - Electrochemical detection of proteins using labeled probes

[0040] Reagents and Devices

[0041] HIV-1 reverse transcriptase (200U, T3610Y, RT) was purchased from GE Healthcare (Quebec, Canada). HIV integrase (100 μg, HIV-122, IN) was purchased from ProspecBio (USA). Bovine serum albumin (BSA), HIV-1 protease (PR) and its inhibitor peptide pepsin (VVStaASta) were purchased from Sigma-Aldrich (Canada). The peptide ligand for RT (VEAIIRILQQLLFIH) (SEQ ID NO: 1) and the peptide ligand for IN (YQLLIRMIYKNI) (SEQ ID No: 2) were purchased from BioBasic (ON, Canada).

[0042] The synthesis of lipoic acid-modified ferrocene (Thc-Fc) for surface modification was carried out as described by Mahmoud et al. (Chem. Eur. J, 2007, 13, 5885-5895), the relevant The content is incorporated herein by reference. Gold microelectrodes (25 μm.d.) were fabricated in the laboratory using well known techniques such as Mahmoud et al.2007. Covalent reagents 1-ethyl-3-(3-dimethy...

Embodiment 2

[0062] Example 2 - Electrochemical detection of proteins using unlabeled probes

[0063] A comparative experiment was performed as follows. The electrochemical detection method of the present invention is performed using unlabeled electrodes and electrodes labeled with molecules that can undergo a reversible one-electron redox process, such as metallocenes, such as ferrocene (Fc). Figure 8 Labeled and unlabeled versions of the detection method of the invention are shown.

[0064] Materials and Reagents

[0065] The working gold electrode is 1.6-mm in diameter and has a geometric area of ​​0.02 cm 2 99.99% (w / w) polycrystalline from CH Instrument Inc. and cleaned before use. Sputtered gold electrodes were prepared by evaporating 200 nm of gold on a silicon wafer with 2 nm of chromium as an adhesion layer. HIV-1 reverse transcriptase enzyme was purchased from Applied Biosystems (Streetsville, ON, Canada). RT-specific peptide (VEAIIRILQQLLFIH) was purchased from Bio Basic...

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PUM

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Abstract

An electrochemical method of identifying the presence of a target protein in a sample is provided. The method comprises providing a redox probe modified to include a detector that is suitable to bind to the target protein, and exposing the sample to the detector-modified redox probe. A change in the electrochemical signal produced by the redox probe as compared to a control signal is indicative of the presence of the target protein.

Description

technical field [0001] The invention relates to a novel electrochemical detection method and a novel probe used in the electrochemical detection. Background technique [0002] Current treatments for AIDS include highly active antiretroviral therapy, abbreviated HAART. Antiretroviral drugs target some essential HIV proteins and inhibit their function. First, fusion inhibitors are designed to terminate HIV entry into host cells. Once the virus enters the cell, the viral RNA is copied into double-stranded cDNA by the viral reverse transcriptase (RT) and then integrated into the host genome. RT has two essential enzymatic activities: DNA polymerization and cleaving of RNA strands in RNA / DNA hybrids (RNase H activity). Anti-RT drugs fall into two classes, both of which target aggregation activity. Most of these drugs are nucleoside analog RT inhibitors (NRTIs), which lack a 3'-OH and thus terminate the DNA chain elongation process upon integration into the replicating strand....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N27/30G01N27/327G01N27/416G01N33/483G01N33/53G01N33/569G01N33/573
CPCG01N2333/16C12Q1/004Y10T29/49716
Inventor H-B·克拉茨K·克曼
Owner UNIV OF WESTERN ONTARIO