A kind of serum albumin with metal chelating function, its preparation method and its application in inhibiting the aggregation of β-amyloid protein
A serum albumin and amyloid technology, applied in the field of biomedicine, can solve the problems of insufficient Aβ aggregation inhibition effect, adsorption inhibition of Aβ aggregation, limited metal ion binding ability, etc. Gentle and easy effect
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Embodiment 1
[0042] Example 1: Synthesis and characterization of metal chelating functional serum albumin with a molar ratio of iminodiacetic acid to serum albumin of 5
[0043] Weigh 20 mg of serum albumin (HSA), dissolve it in 20 mL of boric acid-borax buffer (20 mmol / L, pH 8.0) to obtain a serum albumin solution with a concentration of 1 mg / mL, and add 55 μL of 1,4-butanediol diglycidyl ether. The above mixture was stirred and reacted at 120rpm and 25°C for 8h. After the reaction, the free 1,4-butanediol diglycidyl ether in the system was removed through a SephadexG-25 gel filtration column, and the mobile phase was boric acid-borax buffer (20mmol / L, pH 8.0). Weigh 0.2 mg of iminodiacetic acid powder and add it to the solution obtained in the previous step. The above mixture was stirred and reacted at 120 rpm and 25° C. for 24 h. After the reaction, the free iminodiacetic acid in the system was removed through a SephadexG-25 gel filtration chromatographic column, and the mobile phase...
Embodiment 2
[0049] Example 2: Synthesis and characterization of metal chelating functional serum albumin with a molar ratio of iminodiacetic acid to serum albumin of 50
[0050] 200 mg of serum albumin was weighed and dissolved in 20 mL of boric acid-borax buffer (20 mmol / L, pH 8.5) to obtain a serum albumin solution with a concentration of 10 mg / mL. , and add 825 μL of 1,4-butanediol diglycidyl ether. The above mixture was stirred and reacted at 120rpm and 37°C for 10h. After the reaction, the free 1,4-butanediol diglycidyl ether in the system was removed by SephadexG-25 gel filtration chromatography column, and the mobile phase was boric acid-borax buffer (20mmol / L, pH 8.5). Weigh 19.8 mg of iminodiacetic acid powder and add it to the solution obtained in the previous step. The above mixture was stirred and reacted at 120 rpm and 37° C. for 36 h. After the reaction, remove free iminodiacetic acid in the system through a SephadexG-25 gel filtration chromatographic column, the mobile p...
Embodiment 3
[0056] Example 3: Synthesis and characterization of acidified serum albumin with a molar ratio of iminodiacetic acid to serum albumin of 400
[0057] Weigh 400 mg of serum albumin, dissolve it in 20 mL of boric acid-borax buffer (20 mmol / L, pH 9.0) to obtain a serum albumin solution with a concentration of 20 mg / mL, and add 2200 μL of 1,4-butanediol disulfide glyceryl ether. The above mixture was stirred and reacted at 120rpm and 50°C for 12h. After the reaction, the free 1,4-butanediol diglycidyl ether in the system was removed by SephadexG-25 gel filtration chromatography column, and the mobile phase was boric acid-borax buffer solution (20mmol / L, pH 9.0). Weigh 316.8 mg of iminodiacetic acid powder and add it to the solution obtained in the previous step. The above mixture was stirred and reacted at 120rpm and 50°C for 48h. After the reaction, the free iminodiacetic acid in the system was removed by SephadexG-25 gel filtration chromatographic column, the mobile phase was w...
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