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A kind of mercury-igm chelate and its preparation method and application
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A chelate and chelation reaction technology, applied in the field of detection, can solve problems such as mercury pollution, and achieve the effect of improving accuracy and repeatability
Inactive Publication Date: 2018-01-05
广州锦海骢健康科技有限公司
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[0006] For the serious problem of mercury pollution, the object of the present invention is to provide a kind of mercury-IgM chelate and preparation method thereof, and establish the qualitative and quantitative detection method of mercury-IgM chelate, so that quantitative detection mercury-IgM chelate Application in evaluating the degree of mercury pollution in an area
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[0059] The present invention also provides a preparation method of mercury-IgM chelate, including the following steps:
[0060] A) The chelation reaction of mercury and IgM: mercury ions are added to human-derived IgM or IgM reconstituted according to biological methods to perform a chelation reaction to obtain a reaction solution;
[0061] B) Extraction of purified mercury-IgM chelate: using immunoaffinity chromatography to remove unreacted IgM and excess mercury ions in the reaction solution to obtain mercury-IgM chelate, the specific steps are as follows:
[0062] (1) Dissolve the sample: add physiological saline to the reaction solution obtained in step A) to reconstitute the mercury-IgM chelate
[0063] (2) Equilibrate the chromatography column: use the dilution buffer to flush the pipeline of the chromatography column, load the filler that can specifically bind to IgM in the chromatography column, and continue to use the dilution buffer to balance the chromatography column after ...
Embodiment 1
[0157] Example 1 :The preparation method of mercury-IgM chelate includes the following steps:
[0158] A) The chelating reaction of mercury and IgM: adding mercury ions to the human-sourced IgM to perform the chelating reaction to obtain a reaction solution;
[0159] Reagent preparation:
[0160] 1) Borate buffer (0.01M): Weigh 0.31g of boric acid and dissolve in 400ml of ultrapure water, adjust the pH to 9.0 with 0.1M NaOH, and dilute to 500mL.
[0161] 2) IgM solution: Weigh 4.0mg IgM and dissolve it in 4.0mL 0.01M pH9.0 borate buffer, shake it thoroughly to dissolve, and prepare a 1.0mg / mL protein solution;
[0162] 3) 5mmol / L EDTA+200mmol / LNaHCO 3 Solution: Weigh EDTA·2H 2 O 1.86g, NaHCO 3 Dissolve 16.8g in 900mL ultrapure water, adjust the pH to 8.0 with 1.0M NaOH and dilute the volume to 1000ml, autoclave and store at room temperature;
[0163] 4) ITCBE (purchased from Tongren Chemical Research Institute, Japan, item number M030)
[0164] 5) Dialysis bag (molecular weight cut-off 1...
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Abstract
The invention discloses a mercury-IgM chelate and its preparation method and application. The mercury-IgM chelate is formed by chelating mercury ions and IgM through sulfhydryl groups or / and cysteine residues. The invention establishes a qualitative and quantitative detection method for mercury-IgM chelate, so as to quantitatively detect the application of mercury-IgM chelate in evaluating the degree of mercury pollution in an area. Quantitative detection of mercury-IgM chelate in the serum of a population in an area can indirectly reflect the mercury pollution of the population in this area, thereby indirectly reflecting the degree of mercury pollution in this area. The accuracy of the mercury-IgM chelate quantitative detection method established by the invention is greatly improved, and the repeatability of detection is greatly improved.
Description
Technical field [0001] The invention relates to the field of detection, and more specifically, to a mercury-IgM chelate and a preparation method and application thereof. Background technique [0002] IgM in serum is composed of 5 monomers connected by a J chain and a disulfide bond to form a pentamer, with the largest molecular weight of 970kD, and a sedimentation coefficient of 19S, which is called maHgoglobulin. In terms of molecular structure, IgM has no hinge region, and Cμ2 may replace the function of the hinge region. IgM was the first immunoglobulin to appear in the process of biological evolution. In the process of ontogeny, whether it is the Ig (SmIg) on the surface of the B cell membrane or the Ig synthesized and secreted into the serum, IgM is the first Ig to appear, and the fetus in the late embryonic development is capable of producing IgM. In the process of antigen stimulation to induce humoral immune response, generally IgM is also produced first. IgM accounts...
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