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A kind of chromium-iga chelate and its preparation method and application
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A technology of chelation compound and chelation reaction, which is applied in the detection field, can solve problems such as chromium pollution, and achieve the effect of improving accuracy and repeatability
Active Publication Date: 2018-08-21
上海保亿仁生物医药有限公司
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[0006] For the serious problem of chromium pollution, the object of the present invention is to provide a kind of chromium-IgA chelate and preparation method thereof, and establish the qualitative and quantitative detection method of chromium-IgA chelate, so that quantitative detection chromium-IgA chelate Application in evaluating the degree of chromium pollution in an area
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preparation example Construction
[0058] The present invention also provides a kind of preparation method of chromium-IgA chelate, comprises the following steps:
[0059] A) the chelation reaction of chromium and IgA: add chromium ion to carry out chelation reaction in the IgA of human source, obtain reaction solution;
[0060] B) extraction of purified chromium-IgA chelate: adopt immunoaffinity chromatography to remove unreacted IgA and redundant chromium ions in the reaction solution to obtain chromium-IgA chelate. 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 chromium-IgA 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 IgA, and continue to use the dilution buffer to balance the chromatography column after loading the column; The f...
Embodiment 1
[0144] Example 1 : the preparation method of chromium-IgA chelate, comprises the following steps:
[0145] A) the chelation reaction of chromium and IgA: add chromium ion to carry out chelation reaction in the IgA of human source, obtain 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) IgA solution: Weigh 4.0mg IgA and dissolve it in 4.0mL 0.01M pH9.0 borate buffer, 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 Bi...
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Abstract
The invention discloses a chromium-IgA chelate and its preparation method and application. The chromium-IgA chelate is formed by chelating chromium ions and IgA through sulfhydryl groups or / and cysteine residues. The invention establishes a qualitative and quantitative detection method for the chromium-IgA chelate, so as to quantitatively detect the application of the chromium-IgA chelate in evaluating the degree of chromium pollution in an area. Quantitative detection of chromium-IgA chelate in the serum of a population in an area can indirectly reflect the chromium pollution of the population in this area, thereby indirectly reflecting the degree of chromium pollution in this area. The accuracy of the quantitative detection method for chromium-IgA chelate established by the invention is greatly improved, and the repeatability of detection is greatly improved.
Description
technical field [0001] The invention relates to the detection field, and more specifically relates to a chromium-IgA chelate, a preparation method and application thereof. Background technique [0002] IgA 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 IgA-producing cells, which are mainly derived from the gastrointestinal tract. In humans, there is also a small amount of IgA from the bone marrow. Humans begin to synthesize IgA 4 to 6 months after birth, and the serum content reaches adult levels at 4 to 12 years old, accounting for about 10% of the total Ig of serotype IgA, with a half-life of about 5 to 6 days. IgA has two subclasses, IgA1 and IgA2. IgA1 mainly exists in serum, accounting for about 85% of IgA in ser...
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