Enzyme linked immunosorbent assay kit for detecting content of heavy metal zinc ions in sample

A technology of enzyme-linked immunosorbent reagents and zinc ions, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to use on-site detection, unfavorable promotion and application, and long detection time, so as to achieve easy promotion, easy promotion and application, and time-effective strong effect

Active Publication Date: 2014-01-22
HENAN INST OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional heavy metal ion detection methods such as flameless atomic absorption spectrometry, flame atomic absorption spectrometry, inductively coupled plasma mass spectrometry, voltammetry and ion chromatography, as well as electrothermal atomization atomic absorption spectrometry, must be detected in a large Analytical instruments are carried out in key laboratories, which cannot be used for on-site testing, and are limited by high costs, limited processing capacity, and long testing time, which are not conducive to popularization and application in production, and are difficult to adapt to the environment and on-site spot checks of market products and products. Requirements for rapid customs clearance of imports and exports; immunological detection methods have the advantages of fast, cheap, sensitive and specificity, and can complete the detection of high-throughput samples at the same time. Establishing an immunoassay method to detect zinc ions is a need for production and economic development

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  • Enzyme linked immunosorbent assay kit for detecting content of heavy metal zinc ions in sample
  • Enzyme linked immunosorbent assay kit for detecting content of heavy metal zinc ions in sample
  • Enzyme linked immunosorbent assay kit for detecting content of heavy metal zinc ions in sample

Examples

Experimental program
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Embodiment 1

[0026] Embodiment 1, the preparation of zinc ion chelate, polyclonal, monoclonal antibody

[0027] 1.1 Synthesis and identification of zinc ion chelate

[0028] Weigh 100 mg of zinc chloride with a purity of 99.99% and dissolve it in 100 μL of high-grade concentrated nitric acid. After fully dissolving, add ultrapure water to make the final volume 1 mL to form a zinc solution with a concentration of 733.7 mmoL / L. Weigh 10mg EDTA and dissolve it in 10mM HBS buffer to prepare a 34.2mmoL / L EDTA solution. After mixing the two, adjust the pH value to 6.0 with NaOH, and react on a shaker at room temperature for 24 hours to form Zn 2+ - EDTA chelate solution.

[0029] 100μg / mL Zn 2+ The standard stock solution is diluted with 2% nitric acid to form a concentration gradient of 0 μg / mL, 0.25 μg / mL, 0.5 μg / mL, 1 μg / mL, 2 μg / mL, and 4 μg / mL. The instrument software automatically draws a standard curve and obtains a linear Regression equation; the sample solution was diluted 50 times,...

Embodiment 2

[0038] Embodiment 2, the preparation of zinc ion detection ELISA kit

[0039] 2.1 ELISA kit detection reaction principle

[0040] Immunological detection is based on the principle of antigen and antibody reaction, using known antigens to detect unknown antibodies or using known antibodies to detect unknown antigens. The enzyme-linked immunosorbent assay kit selects the appropriate concentration of the coated antigen to coat the enzyme-labeled plate, takes the processed sample to be tested as required and reacts with the polyclonal or monoclonal antibody provided, and the antibody that is not fully combined with the enzyme label The sites on the plate are combined, through the binding reaction of the goat anti-mouse or rabbit anti-mouse secondary antibody labeled with horseradish peroxidase, the color development is reflected by the color development, and the results are observed, or the degree is calculated with a microplate reader, and calculated according to the results. Th...

Embodiment 3

[0056] Embodiment 3, the performance of zinc ion ELISA kit

[0057] 3.1 Sensitivity determination according to the minimum detection limit of blocking ELISA is B / B 0 %=120%, calculate the sensitivity of the kit according to the curve regression equation, and determine the detection limit. As a result, the detection limit was 0.248μg / L. According to the results obtained by blocking ELISA, B / B 0 %=10%~90% is the detection range, according to the curve regression equation, the detection range of the kit is 3.774~442.39μg / L.

[0058] 3.2 Determination of accuracy will Zn 2+ Standards were added to soil, tap water and pork at final concentrations of 5, 10, 20 and 40 μg / L, respectively, and 6 replicates were set up, and the accuracy was determined by recovery and coefficient of variation (CV). The results are shown in Table 2. The recovery rate of soil is 84.15%-87.6%, the average is 85.36%, and the coefficient of variation (CV) is 6.05%-13.09%; the recovery rate of tap water i...

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Abstract

The invention provides an enzyme linked immunosorbent assay kit for detecting content of heavy metal zinc ions in a sample, and the enzyme linked immunosorbent assay kit comprises an elisa plate with a coating antigen, an enzyme marker, a zinc ion chelate specific antibody, a zinc ion chelate standard substance solution, a substrate coloured solution, a stop solution, a washing concentrate and a concentrated complex solution. The invention also provides a method for detecting a zinc ion chelate by the enzyme linked immunosorbent assay kit, and the method comprises the following steps: firstly, carrying out sample pretreatment; then carrying out detection by the kit; and finally, analyzing a detection result. The invention provides the enzyme linked immunosorbent assay kit and the detection method for detecting the residual quantity of the zinc ions in the samples such as foods, feeds, beverages, soil, animal tissues and the like; the operation is simple and convenient; the cost is low; the flexibility is high; and the enzyme linked immunosorbent assay kit can be used for field monitoring and is suitable for screening a great amount of samples.

Description

Technical field: [0001] The invention relates to a monoclonal antibody and an ELISA kit for detecting heavy metal ion zinc ions. Background technique: [0002] Heavy metals in soil and agricultural products are closely related to human health. Heavy metal pollution mainly refers to mercury, chromium, cadmium, lead, and metalloid arsenic with significant biological toxicity, and also includes toxic heavy metals such as copper, cobalt, nickel, and tin. With the expansion of cities and the development of large-scale industries, after a large amount of heavy metals enter the environment, even if the concentration is very low, they may cause harm. They will eventually threaten human health through drinking water, or through bioaccumulation and food chains. Heavy metal pollution is different from other types of pollution in that it is concealed, long-term and irreversible. [0003] Among the metals used by contemporary humans, zinc (Zn) is the fourth largest heavy metal after ir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/566
CPCG01N33/577
Inventor 张海棠王爱萍王自良范国英姜金庆王顺岗
Owner HENAN INST OF SCI & TECH
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