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

An enzyme-linked immunosorbent reagent and copper ion technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that it is difficult to adapt to the requirements of rapid customs clearance of products, and cannot use on-site detection, etc., to achieve easy promotion, easy promotion and application, and judgment image effect

Active Publication Date: 2014-02-05
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditional heavy metal monitoring methods such as atomic absorption spectrometry and inductively coupled plasma emission spectrometry require large-scale analytical instruments, which cannot be used for on-site testing, and are difficult to adapt to the requirements of the environment, on-site spot checks of agricultural and livestock products, and rapid customs clearance of product imports and exports.

Method used

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

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Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1, the preparation of copper ion chelate, polyclonal, monoclonal antibody

[0026] 1.1 Transformation and identification of copper ion chelate

[0027]Weigh 76.7 mg of copper nitrate 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 copper solution with a concentration of 409 mmoL / L. Weigh 10mg EDTA and dissolve in 10mM HBS buffer to prepare EDTA solution. After mixing the two evenly, adjust the pH value to 6.0 with NaOH, and react on a shaker at room temperature for 24 hours to form Cu 2+ - EDTA chelate solution.

[0028] 100 μg / mL of Cu 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 t...

Embodiment 2

[0036] Embodiment 2, preparation of copper ion detection ELISA kit

[0037] 2.1 detection reaction principle of immunoassay kit

[0038] The immunoassay kit selects the appropriate coating antigen concentration to coat the microtiter plate, and the detection kit made after blocking with heterologous protein can be used for the detection of samples. Take the treated samples to be tested and react with the provided polyclonal or monoclonal antibodies as required. The incompletely bound antibodies bind to the sites on the microtiter plate, and the goat antibody labeled with horseradish peroxidase Mouse or rabbit anti-mouse secondary antibody binding reaction, through color development to reflect the color development, observe the results, or use a microplate reader to read the degree, and calculate the content of the analyte according to the results. The judgment of the result is compared with the provided negative and positive controls. If the color depth is exactly the same as...

Embodiment 3

[0052] Embodiment 3, the performance of copper ion ELISA kit

[0053] 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 0.42 μg / 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 2.83~456.04μg / L.

[0054] 3.2 Accuracy determination of Cu 2+Standards were added to soil and tap water at final concentrations of 5, 10, 20, and 40 μg / L, respectively, and six replicates were set up. The accuracy was determined by recovery and coefficient of variation (CV). The results are shown in Table 2. The recovery rate of soil is 77.25%-82.3%, with an average of 80.56%, and the coefficient of variation (CV) is 9.5%-13.7%; the recovery rate of tap water is 80.8%-88...

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Abstract

The invention discloses an enzyme linked immunosorbent assay kit used for detecting the content of heavy metal copper ions in a sample. The kit includes a coat antigen coated enzyme label plate, an enzyme marker, a copper ion chelate specific monoclonal antibody or polyclonal antibody, a copper ion chelate standard solution, a substrate color development solution, a terminating solution, a lotion, and a diluted chelating agent EDTA. The enzyme linked immunosorbent assay kit used for copper ion detection has the advantages of: (1) strong specificity, high sensitivity, a lowest detection limit of 0.42 microgram / L, and a detection range of 2.83-456.04 microgram / L; (2) simplicity, rapidity, and strong timeliness; (3) vivid, visual and accurate result display; and (4) low cost, wide application range, convenient popularization, on-site supervision and suitability for screening a large number of samples.

Description

Technical field: [0001] The invention relates to a monoclonal antibody and an ELISA kit for detecting copper ions. Background technique: [0002] 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] Heavy metal copper is an important metal element existing in nature. Copper and its compounds are widely used in industry and agriculture due to their various excellent properties. Constant coppe...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/566
CPCG01N33/531G01N33/566G01N33/577G01N33/683
Inventor 范国英王自良王爱萍张海棠姜金庆王顺岗
Owner HENAN INST OF SCI & TECH
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