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Method for detecting microcystin-LR by self-assembly based on end face of gold nano-rod

A technology of microcystin and gold nanorods, which is applied in the field of immunoassay chemistry, can solve the problems that cannot meet the requirements of fast, convenient and accurate, poor specificity, and no commercial supply of specific protein phosphatase, and achieve the goal of simplifying the reaction Conditions, reduce labor intensity, improve the effect of sensitivity

Inactive Publication Date: 2012-09-26
JIANGNAN UNIV
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Problems solved by technology

Instrumental analysis methods mainly include high-performance liquid chromatography (HPLC), mass spectrometry, liquid chromatography-mass spectrometry, etc. Although these methods are sensitive, they require expensive instruments and equipment, professional operators, and relatively high requirements for inspection materials. Further sample pretreatment can be carried out, which can no longer meet the requirements of modern detection for fast, convenient and accurate
The biochemical method is mainly the protein phosphatase inhibition test method, which has the advantage of being fast and can detect a large number of samples in a few hours, but its biggest disadvantage is that specific protein phosphatases are not commercially available and have poor specificity

Method used

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  • Method for detecting microcystin-LR by self-assembly based on end face of gold nano-rod
  • Method for detecting microcystin-LR by self-assembly based on end face of gold nano-rod
  • Method for detecting microcystin-LR by self-assembly based on end face of gold nano-rod

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Embodiment 1

[0048] (1) Coating original modification:

[0049] The modification of the coating source uses the carbodiimide method to combine lipoic acid and the coating source. The specific operation steps are as follows:

[0050] ① 0.3 mL of lipoic acid ethanol solution (containing 1.625 mg of lipoic acid) was mixed with 0.2 mL of the original coating solution.

[0051] ② Add 75 μL of carbodiimide-containing dropwise into the above solution under the condition of magnetic stirring, and react at room temperature for 4 hours.

[0052] ③Dialyze with pH 7.4 phosphate buffered saline for 3 days, changing the solution 3 times a day.

[0053] ④ Store the modified coating at 4°C for later use.

[0054] (2) Antibody modification:

[0055] ① Mix 0.3mL of lipoic acid-containing ethanol solution (containing 1.625mg of lipoic acid) with 0.4mL of antibody.

[0056] ②Put 75 μL of carbodiimide dropwise into the above solution under magnetic stirring, and react at room temperature for 4 hours.

[0...

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Abstract

The invention discloses a method for detecting microcystin-LR by self-assembly based on the end face of a gold nano-rod, belonging to the technical field of chemical immunoassay. In the invention, the end face of the gold nano-rod is respectively coupled with microcystin coating antigen and anti-microcystin antibody to form a probe of the gold nano-rod; and the synthesized probe of the gold nano-rod performs immunological reaction with the microcystin-LR so as to achieve the detection of the microcystin-LR through the change of the grain diameter of a nano-chain which is formed by the antigenantibody reaction of the end face of the gold nano-rod. The invention performs reaction in a liquid environment, only one step of reaction is needed without washing step, which simplifies the condition of the reaction, reduces labour intensity and improves the detection sensitivity.

Description

technical field [0001] The invention discloses a method for detecting microcystin-LR (MC-LR) based on the end face self-assembly of gold nanorods, which belongs to the technical field of immunoanalysis chemistry. Background technique [0002] With the development of the economy, sewage containing a large amount of nitrogen and phosphorus nutrients enters lakes and reservoirs to eutrophicate water bodies, resulting in abnormal proliferation of algae and the release of secondary metabolites algae toxins (Algae Toxins), threatening the safety of human drinking water and the safety of other organisms in the water. Among them, Microcystins (MCs) are the most widely distributed and most harmful, and their structures are as follows: [0003] [0004] It contains five fixed amino acids and two variable amino acids X, Y. That is, in the above structural formula, X and Y are Leu and Arg respectively, which is MC-LR. MC-LR is the most common and most toxic microcystin. The maximu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/531G01N15/02C07K16/14
Inventor 胥传来徐丽广陈伟朱颖越刘丽强徐乃丰马伟许定华
Owner JIANGNAN UNIV
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