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A kind of mercury-ige 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: 2018-02-16
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-IgE chelate and preparation method thereof, and establish the qualitative and quantitative detection method of mercury-IgE chelate, so that quantitative detection mercury-IgE chelate Application in evaluating the degree of mercury pollution in an area
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preparation example Construction
[0058] The present invention also provides a kind of preparation method of mercury-IgE chelate, comprises the following steps:
[0059] A) chelation reaction of mercury and IgE: adding mercury ions to human-sourced IgE for chelation reaction to obtain a reaction solution;
[0060] B) Extraction of purified mercury-IgE chelates: Immunoaffinity chromatography is used to remove unreacted IgE and redundant mercury ions in the reaction solution to obtain mercury-IgE chelates. The specific steps are as follows:
[0061] (1) Dissolving the sample: adding physiological saline to the reaction solution obtained through step A) to redissolve the mercury-IgE chelate;
[0062] (2) Equilibrium chromatography column: use the dilution buffer to flush the pipeline of the chromatography column, fill the chromatography column with a filler that can specifically bind to IgE, and continue to use the dilution buffer to balance the chromatography column after loading the column; The filler capable ...
Embodiment 1
[0144] Example 1 : the preparation method of mercury-IgE chelate, comprises the following steps:
[0145] A) chelation reaction of mercury and IgE: adding mercury ions to human-sourced IgE for chelation reaction to obtain a reaction solution;
[0146] Reagent preparation:
[0147] 1) Borate buffer (0.01M): Weigh 0.31g of boric acid and dissolve it in 400mL of ultrapure water, adjust the pH to 9.0 with 0.1M NaOH, and dilute to 500mL.
[0148] 2) IgE solution: Weigh 4.0mg IgE and dissolve it in 4.0mL 0.01M pH9.0 borate buffer solution, fully shake and dissolve, and prepare a 1.0mg / mL protein solution;
[0149] 3) EDTA-NaHCO3 mixed 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 to 1000mL, autoclave, and store at room temperature;
[0150] 4) ITCBE was purchased from Japan Tongren Chemical Research Institute, the article number is M030;
[0151] 5) The dialysis bag was purchased from Biosho...
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Abstract
The invention discloses a mercury-IgE chelate and its preparation method and application. The mercury-IgE chelate is formed by chelating mercury ions and IgE through sulfhydryl groups or / and cysteine residues. The invention establishes a qualitative and quantitative detection method for mercury-IgE chelate, so as to quantitatively detect the application of mercury-IgE chelate in evaluating the degree of mercury pollution in an area. Quantitative detection of mercury-IgE 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 mercury-IgE chelate quantitative detection method established by the invention has greatly improved accuracy and greatly improved detection repeatability.
Description
technical field [0001] The invention relates to the field of detection, and more specifically relates to a mercury-IgE chelate and its preparation method and application. Background technique [0002] IgE is mainly produced by mucosa-associated lymphoid tissues, most of which are synthesized by gastrointestinal lymphoid tissues, and a small part is synthesized by mucosal tissues of the respiratory tract, salivary glands and reproductive tract. Lactating maternal glandular tissue contains a large number of IgE-producing cells, which are mainly derived from the gastrointestinal tract. In humans, there is also a small amount of IgE from the bone marrow. Humans begin to synthesize IgE 4-6 months after birth, and the content in serum reaches adult levels at 4-12 years old, accounting for about 10% of the total Ig of serotype IgE, with a half-life of about 5-6 days. IgE has two subclasses, IgE1 and IgE2. IgE1 mainly exists in serum, accounting for about 85% of IgE in serum, and...
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