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In-situ anode dissolving-out volt-ampere analytical method based on metal marking and biology affinity

An anodic stripping voltammetry and metal labeling technology, which is applied in the direction of analyzing materials, material analysis by electromagnetic means, and measuring devices, can solve the problems of increasing the complexity of experimental operations, limiting the detection sensitivity of metal immunoelectric analysis, and improving the The effect of electrochemical signal output, enrichment efficiency improvement, and simplified operation steps

Active Publication Date: 2013-12-25
HUNAN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the past, the strategy of dissolving metal markers and then transferring the solution for voltammetric analysis was usually used in the past, so the inevitable dilution of the solution limited the detection sensitivity of metal immunoelectroanalysis, and the replacement of the solution also increased the experimental operation. complexity

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  • In-situ anode dissolving-out volt-ampere analytical method based on metal marking and biology affinity
  • In-situ anode dissolving-out volt-ampere analytical method based on metal marking and biology affinity
  • In-situ anode dissolving-out volt-ampere analytical method based on metal marking and biology affinity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Gold nanoparticles (AuNPs)-labeled secondary antibodies, CdS quantum dots (CdS QDs)-labeled secondary antibodies (Ab 2 -CdS), immune electrodes were prepared according to the following method:

[0037] (1) Preparation of AuNPs-labeled secondary antibody (Ab 2 -AuNPs): In 100 mL of boiling water, under vigorous stirring, add 250 mL of 4% (mass concentration) HAuCl 4 , then add 2.5 mL of 1% (mass concentration) sodium citrate dropwise, and heat for another 15 min when the solution turns wine red. After heating was stopped, the mixture was cooled to room temperature with stirring. Add 30 mg of secondary antibody (Ab 2 ), adsorbed overnight. After centrifugation at 4800 rpm for 30 min at low speed, the cells were washed twice with phosphate buffered saline (PBS). Finally, the conjugate was redispersed in 1 mL 0.1 mol / L PBS containing 1% bovine serum albumin (BSA) to increase the stability of the immunogold gel and reduce the non-specific adsorption in the analysis. Ab...

Embodiment 2

[0061] Example 2: Electroanalysis of nucleic acid aptamers based on the method of the present invention

[0062] This example is based on the electroanalysis of nucleic acid aptamer affinity, metal labeling and anodic stripping voltammetry to detect thrombin, using the technical solution of the present invention to obtain the response in the nucleic acid aptamer electrode.

[0063] This embodiment includes the following steps:

[0064] Sample determination:

[0065] 1. Nucleic acid aptamer analysis by gold-labeled silver staining:

[0066] The method of the present invention: the implementation steps of the "method of the present invention" in the "gold standard silver staining immunoassay" in Example 1 are the same.

[0067] 2. Analysis of nucleic acid aptamers labeled with CdS QDs:

[0068] The method of the present invention: the implementation steps of the "method of the present invention" in the "immunoassay of CdS QDs labeling" in Example 1 are the same.

[0069] Ana...

Embodiment 3

[0070] Embodiment 3: The method of the present invention based on the gold-labeled silver-stained immunoelectrode set for simultaneous detection of carcinoembryonic antigen (hCEA) and alpha-fetoprotein (hAFP) two target analytes

[0071] Sample determination: the implementation steps of the "method of the present invention" in the "gold standard silver staining immunoassay" in Example 1 are the same. Add release agent 5 mL 1 mol / L HNO to test conditions 3 .

[0072] Analysis of measurement results: discuss in conjunction with Figure 5. Fig. 5 shows the signal response standard curve of the gold-labeled immune electrode for detecting different concentrations of antigens based on the anodic stripping voltammetry of the electrode group according to the present invention. It can be seen from the figure that as the antigen concentration increases, the current response also increases. The linear range of hCEA and hAFP is from 4 fg / mL to 400 ng / mL, and the lower detection limits ar...

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Abstract

An in-situ anode dissolving-out volt-ampere analytical method based on metal marking and biology affinity comprises the following steps that (1) an immune electrode or a nucleic acid aptamer electrode is used as a working electrode, an electrochemistry instrument is connected in air, cathode potential which can achieve metal marker metal ion electro-deposition is exerted in advance, then a small-size releasing agent is added to the surface of the working electrode, an electrolytic tank is connected, so that a metal marker is dissolved, metal ions are released and are continuously and electrically reduced to atom state metal, and accordingly a metal layer is formed in the surface of the working electrode in a gathering mode; (2) anode dissolving-out current signals of the metal layer gathered on the surface of the working electrode are directly obtained on the surface of an original working electrode, and accordingly quantitative analysis on target analyte in a sample is achieved indirectly. The in-situ anode dissolving-out volt-ampere analytical method can be used for single-target analyte detecting and multi-target analyte multi-channel detecting based on metal marking and biology affinity.

Description

technical field [0001] The present invention relates to an electrochemical quantitative analysis method based on metal labeling and bioaffinity, in particular, relates to a method based on bioaffinity (such as immune, nucleic acid aptamer and other bioaffinity types), metal labeling and anode Stripping voltammetry is a method for quantitative analysis of target analytes in samples. Background technique [0002] In recent years, bioanalysis techniques based on bioaffinity principles, such as immunoassay and nucleic acid aptamer analysis, have developed rapidly and have been widely used in fields such as health and environmental protection [D. W. Kimmel, G. LeBlanc, M. E. Meschievitz, and D. E. Cliffel, Anal. Chem. 2012, 84, 685-707]. It can be considered that for the immunoassay method of the designated immune pair, the immune system has excellent specific recognition characteristics and presents excellent selectivity, so improving the sensitivity of the immunoassay has beco...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N27/327
Inventor 谢青季覃晓丽傅迎春陈超谭月明黄毅部丽娟
Owner HUNAN NORMAL UNIVERSITY
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