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Preparation method of immunomagnetic beads for detecting edetate, and prepared immunomagnetic beads

A technology of EDTA and disodium EDTA, applied in the field of chemical detection, can solve the problem of excluding the detection of EDTA, the absence of disodium EDTA, and the detection results. Large error and other problems, to achieve the effect of less impurities, good protection and stable magnetic beads, and improved accuracy

Pending Publication Date: 2020-01-14
HENAN BUSINESS SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods of EDTA in food include gas chromatography, high performance liquid chromatography and liquid chromatography tandem mass spectrometry; standard methods include GB5009.278-2016 and SN / T 3855-2014, but None of the above detection methods include the detection of EDTA in flour and flour products
There is currently no reference standard for the detection of disodium edetate in flour and flour products
Moreover, the above-mentioned disodium edetate detection method has complex operation steps, time-consuming, poor specificity, and low detection sensitivity; especially when EDTA-Na 2 When the content is low, the error of the detection result is large and the accuracy is low

Method used

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  • Preparation method of immunomagnetic beads for detecting edetate, and prepared immunomagnetic beads
  • Preparation method of immunomagnetic beads for detecting edetate, and prepared immunomagnetic beads
  • Preparation method of immunomagnetic beads for detecting edetate, and prepared immunomagnetic beads

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Screening of buffer

[0033] Take four parts of 200 μL magnetic beads, place them in four centrifuge tubes, shake for 1 min and place them on a magnetic stand; add 400 μL of ddH to the four centrifuge tubes respectively 2 O, PBS, Earle's BSS solution (pH 7.0), carbonate buffer solution, and then add 200 μL EDTA monoclonal antibody solution with a concentration of 0.1 mg / mL to each centrifuge tube, shake and mix well , and then put the four centrifuge tubes on a shaker at 30°C for 4 hours, put them on a magnetic stand to stand for stratification, separate the supernatant, and measure the disodium ethylenediaminetetraacetic acid monosodium in the supernatant with a UV spectrophotometer. Clonal antibody concentration. Wherein, the Earle'sBSS solution is made of NaCl, KCl, Na 2 HPO 4 、KH 2 PO 4 、CaCL 2 , MgSO 4 , glucose, NaHCO 3 , Tween-20 and water, the concentration of each component in Earle’s BSS solution is: NaCl is 6.8g / L, KCl is 0.4g / L, NaCl is...

Embodiment 2

[0037] Embodiment 2: Screening of buffer pH

[0038] Take four parts of 200 μL magnetic beads, place them in four centrifuge tubes, shake for 1 min, place them on a magnetic stand, add 400 μL of Earle’s BSS solutions with pHs of 6.8, 7.0, 7.4, and 8.0 to the four centrifuge tubes, and then separately Add 200 μL of disodium edetate monoclonal antibody with a concentration of 0.1 mg / mL to each centrifuge tube, shake and mix evenly, then place the four centrifuge tubes on a shaker at 30°C for 4 hours, and place them on a magnetic stand Leave to stand and separate layers, separate the supernatant, and measure the monoclonal antibody concentration in the supernatant with an ultraviolet spectrophotometer. Among them, Earle's BSS solution is made of NaCl, KCl, Na 2 HPO 4 、KH 2 PO 4 、CaCL 2 , MgSO 4 , glucose, NaHCO 3 , Tween-20 and the mixed solution that water forms, the concentration of each component in the Earle's BSS solution is identical with embodiment 1. The experimen...

Embodiment 3

[0042] Example 3: Screening of Coupling Activation Reaction Temperature

[0043] Take six parts of 200 μL magnetic beads, place them in six centrifuge tubes, shake for 1 min and place them on a magnetic stand, add 400 μL of Earle’s BSS solution with pH 7.0 to each of the six centrifuge tubes, and then add to each Add 200 μL of EDTA disodium monoclonal antibody with a concentration of 0.1 mg / mL into the centrifuge tube, shake and mix evenly, and then place the six centrifuge tubes at 10°C, 20°C, 25°C, 30°C, and 37°C respectively and react at 45°C. After 2 hours of reaction, place the centrifuge tubes on a magnetic stand to stand for stratification, separate the supernatant, and measure the concentration of EDTA monoclonal antibody in the supernatant with a UV spectrophotometer . Wherein, said Earle's BSS solution is made of NaCl, KCl, Na 2 HPO 4 、KH 2 PO 4 、CaCL 2 , MgSO 4 , glucose, NaHCO 3 , Tween-20 and water, the concentration of each component in the Earle’s BSS so...

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Abstract

The invention belongs to the technical field of chemical detection, and discloses a preparation method of immunomagnetic beads for detecting edetate, and prepared immunomagnetic beads. The preparationmethod of the immunomagnetic beads is as follows: (1) taking magnetic beads, suspending the magnetic beads in a buffer solution after washing the same; (2) adding an edetate monoclonal antibody solution into the magnetic bead suspension, performing oscillation and uniform mixing, performing coupling activation at 10-45 DEG C for 20-120min, then performing stewing and layering on a magnetic forceframe, and discarding the supernatant to obtain a magnetic bead-antibody compound; and (3) adding the magnetic bead-antibody compound into the buffer solution to suspend the same, adding confining liquid, performing incubation at room temperature, and performing stewing and layering on the magnetic force frame to obtain the immunomagnetic beads. The immunomagnetic beads prepared by the preparationmethod disclosed by the invention have high specificity and high sensitivity, and can quickly and specifically bind to the edetate in a sample solution to be tested, quickly enrich the edetate in thesample to be tested and greatly improve the detection limit and the detection accuracy of the edetate.

Description

technical field [0001] The invention belongs to the technical field of chemical detection, and in particular relates to a method for preparing immunomagnetic beads used for detecting ethylenediamine tetraacetate and the prepared immunomagnetic beads. Background technique [0002] Disodium ethylenediaminetetraacetic acid (EDTA-Na 2 ) is listed as a food additive allowed to be used in the national food safety standard GB2760-2014 "Usage Standards of Food Additives". In canned and preserved fruit and other foods. Because ethylenediaminetetraacetic acid can form chelates with calcium, magnesium and other trace metals necessary for the human body, long-term excessive intake will lead to the loss of trace metal elements in the human body to a certain extent, causing harm to the human body, and EDTA-Na 2 It will cause irritation to eyes, skin, mucous membranes and respiratory tract. If EDTA-Na is consumed in large quantities for a long time 2 Food that exceeds the standard will ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/577
CPCG01N33/531G01N33/577
Inventor 周莉陈世伟朱海华王晓瑞杜瑞平洋谭静闫格王珂李远远
Owner HENAN BUSINESS SCI RES INST
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