Nickel-IgG chelate as well as preparation method and application thereof
A chelate and chelation reaction technology, applied in the field of detection, can solve problems such as nickel pollution, and achieve the effect of improving repeatability and accuracy
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[0059] The present invention also provides a kind of preparation method of nickel-IgG chelate, comprises the following steps:
[0060] A) the chelation reaction of nickel and IgG: adding nickel ions to human IgG or IgG recombined according to biological methods for chelation reaction to obtain a reaction solution;
[0061] B) Extraction of purified nickel-IgG chelates: Immunoaffinity chromatography is used to remove unreacted IgG and excess nickel ions in the reaction solution to obtain nickel-IgG chelates. The specific steps are as follows:
[0062] (1) Dissolving sample: add physiological saline to the reaction solution obtained through step A) to redissolve the nickel-IgG chelate
[0063] (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 IgG, and continue to use the dilution buffer to balance the chromatography column after loading the ...
Embodiment 1
[0157] Example 1 : the preparation method of nickel-IgG chelate, comprises the following steps:
[0158] A) the chelation reaction of nickel and IgG: adding nickel ions to the IgG of human origin for chelation reaction to obtain a reaction solution;
[0159] Reagent preparation:
[0160] 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.
[0161] 2) IgG solution: Weigh 4.0mg IgG 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;
[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 to 1000ml, autoclave, and store at room temperature;
[0163] 4) ITCBE (purchased from Japan Tongren Chemical Research Institute, item number M030)
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