Immunoassay method based on enzyme-induced sulfite as activator
An immunoassay and sulfite technology, applied in the field of nanomaterials and biological analysis, can solve problems affecting color stability and other issues, and achieve significant economic and social benefits, convenient operation, and high sensitivity
Inactive Publication Date: 2016-03-30
FUZHOU UNIV
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In the process of color development, it is easily affected by external conditions (pH and temperature), which affects the stability of color development
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Embodiment 1
[0029] 1. Preparation of antibody immobilized on magnetic beads
[0030] 50mgFe 3 o 4 The magnetic beads were added to 1 mL of absolute ethanol, and the solution was sonicated at room temperature for 10 minutes. 30 μL of 3-aminopropyltriethoxysilane was added thereto, and stirred at room temperature for 6 hours. Results The solution was magnetically separated, dispersed in 1 mL of phosphate buffer (PBS, pH 7.4, 300 μL of glutaraldehyde), and stirred for 6 hours. After magnetic control separation, the pellet was dissolved in 1 mL of carbonic acid buffer (pH 9.6, 100 μg of prostate-specific antibody) and shaken overnight at 4 °C. Then the excess antibody was separated and removed, 10wt% bovine albumin (BSA)-PBS (1 mL, pH 7.4) was added, and reacted at 4°C for 2 hours. Finally, add 50 µL of sodium cyanoborohydride and incubate at 4 °C for 1 h. The resulting antibody-immobilized magnetic bead pellet was dispersed in 0.5mL PBS (pH7.4, 1.0wt% BSA, 0.1wt% sodium azide).
[0031...
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Abstract
The invention discloses an immunoassay method based on enzyme-induced sulfite as an activator, and belongs to the technical field of nanomaterials and biological analysis. In the method, the glucose oxidase on the enzyme-labeled nano-probe combined with the target object catalyzes glucose to generate hydrogen peroxide, and then sequentially adds nitric acid, sodium sulfite, silver nitrate and TMB color developing solution, and is used for quantitative detection by an ultraviolet-visible spectrophotometer. The purpose of the present invention is to provide an immunoassay method based on enzyme-induced sulfite as an activator, which is mainly used for the detection of low-abundance protein content in serum. The solution is colored due to the enzyme-induced sulfite as an activator. Utilizing this feature, a colorimetric immunoassay method with high sensitivity and stable performance can be established. The method has the advantages of low cost, simple colorimetric detection method, rapid color development and high sensitivity, and provides a sensitive and stable method for clinical immunological detection.
Description
technical field [0001] The invention relates to an immunoassay method based on enzyme-induced sulfite as an activator, and belongs to the technical field of nanometer materials and biological analysis. 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. Generally, the chromogenic reagents used in colorimetric immunoassays are o-phenylenediamine, benzidine and 3,3',5,5'-tetramethylbenzidine. Among them, 3,3',5,5'-tetramethylbenzidine has become the most widely used chromogenic agent because of its non-carcinogenicity, non-mutation, good stability and high sen...
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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 唐点平赖文强庄君阳
Owner FUZHOU UNIV