Method of using nano zeolite molecular sieve assemble material as affinity chromatography filler to separate functional protein molecules
A technology of nano-zeolite and chromatographic filler, which is applied in the field of nano-zeolite molecular sieve assembly material as an affinity chromatographic filler to separate functional protein molecules, can solve the problems of large diffusion resistance, limited use range, narrow pore channels, etc., and achieves uniform pore channels and convenient repeated regeneration. , the effect of stable nature
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example 1
[0040] 0.1 g of self-made modified nano-beta zeolite filler was adjusted to a slurry with an appropriate amount of deionized water, and transferred to a separation column with a bed height of about 1 cm. Then add 1.0ml equilibration buffer solution to equilibrate the column. Dilute 0.50ml to a concentration of 124μgL -1 1.5mgL of histidine or 1.0mL -1 Peptide solution samples (samples are artificially synthesized polypeptide molecules containing different histidine structures) were transferred to the extraction column, and after the liquid flowed out, 1.0ml equilibration buffer solution was added for washing, and the histidine or polypeptide adsorbed on the column passed through Add formic acid-ammonium formate elution buffer solution with pH of 7.0, 6.0, 5.0, and 4.0 in sequence for gradient elution, the elution rate is 10ml / hr, and then detect the absorbance value in each collection tube by ultraviolet spectroscopy, and draw the outflow distribution map. Example 2-5
example 2-5
[0041] Adjust the bed height of the column to be 1.5, 2.0, 2.5, and 3.0 cm respectively, and the other conditions are the same as above, and repeat the above separation experiment. Example 6-8
example 6-8
[0042] Select respectively formic acid-ammonium formate buffer solution system or sodium phosphate-phosphate buffer solution or imidazole gradient solution as the elution system to separate the polypeptide molecule mixture, repeat the example 1 step (the artificially synthesized polypeptide molecules A and B respectively contain different groups amino acid structure, and mixed in a 1:1 molar ratio). The separation of selenoprotein-P in example 9 mouse plasma
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