Immunoassay method based on platinum nanoparticle mimic enzyme

A platinum nanoparticle and immunoassay technology, applied in the field of immunoassay based on platinum nanoparticle-mimicking enzymes, can solve the problems of insufficient immunoassay sensitivity, insufficient catalytic activity, low detection line, etc. The effect of large-scale preparation and low detection line

Inactive Publication Date: 2013-04-24
FUZHOU UNIV
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Problems solved by technology

However, at present, the catalytic activity of these nanomaterials to simulate enzymes is not high enough, and complex modifications are often required before they can be labeled on antibodies.
These deficiencies lead to the fact that the sen

Method used

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  • Immunoassay method based on platinum nanoparticle mimic enzyme
  • Immunoassay method based on platinum nanoparticle mimic enzyme
  • Immunoassay method based on platinum nanoparticle mimic enzyme

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Embodiment 1

[0028] 1. Preparation of Platinum Nanoparticle Mimetic Enzymes

[0029] In the state of stirring, add 400mL potassium chloroplatinate (K 2 PtCl 6 ) solution. After 1 minute, add 100 mL of 1% (w / w) sodium citrate to the above mixture, and then add 400 mL of freshly prepared 1% (w / w) sodium citrate and 0.08% (w / w) a mixed solution of sodium borohydride. The above reaction solution was placed in an oil bath at 90° C. and heated for 10 minutes to obtain a stock solution of platinum nanoparticle-mimicking enzyme. The prepared platinum nanoparticles mimic enzymes were characterized by transmission electron microscopy, such as figure 1 shown. From figure 1 It can be seen that the size distribution of the prepared platinum nanoparticles is uniform, and the particle size distribution is between 3 and 5 nm.

[0030] 2. Preparation of platinum-labeled antibody nanoprobe complexes

[0031] First, 1.0 mL of the prepared platinum nanoparticle mimic enzyme was adjusted to pH 8-9 wit...

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Abstract

The invention discloses an immunoassay method based on a platinum nanoparticle mimic enzyme. The immunoassay method comprises the following steps: step one, preparing the platinum nanoparticle mimic enzyme through a sodium borohydride reduction method; step two, labeling the platinum nanoparticle mimic enzyme onto an antibody, and forming a platinum-labeled antibody nanoprobe; step three, forming an immune complex by the antibody coated on an enzyme-labeled microporous plate, a sample to be detected and the platinum-labeled antibody nanoprobe through a sandwich type immunoassay reaction mode; and step four, adding a chromogenic substrate solution to perform chromogenic detection. The immunoassay method based on the platinum nanoparticle mimic enzyme can be used for the colorimetric immunoassay of a macromolecular object; the related nanoprobe complex is simple in preparation process and low in cost; and the immunoassay method is simple, rapid in color development, and high in sensitivity, thereby providing a novel enzyme-free, sensitive and stable method for the clinical immunological detection.

Description

technical field [0001] The invention belongs to the technical field of nanomaterials and biological analysis, in particular to an immunoassay method based on platinum nanoparticle imitating enzymes. Background technique [0002] Due to its unique specificity, immunoassay technology has been widely used in clinical diagnosis and environmental detection. Since its establishment in 1971, enzyme-linked immunoassay (ELISA) has been rapidly developed and applied due to its advantages of fastness, sensitivity, simplicity, and easy standardization, and has gradually become one of the most widely used immunoassay methods. Involved in various fields of immunological testing. However, if some essential shortcomings of the protease used in it can be overcome, such as protease is easy to inactivate, low stability, etc., then the performance of ELISA may be further improved. In addition, extraction and purification of proteases is often time-consuming, expensive and complicated; prepara...

Claims

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Application Information

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IPC IPC(8): G01N33/545
Inventor 唐点平高壮强杨黄浩陈国南
Owner FUZHOU UNIV
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