Method for extracting various urinary proteins from urine
A urine protein and urine technology, which is applied in the field of protein extraction, can solve the problems of the specific product type and output that cannot be adjusted, the separation and extraction of one type from a batch of urine, and the high production cost.
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Embodiment 1
[0056] Example 1 Optimization of the method of the invention
[0057] (1) Dialysis solution, equilibrium buffer solution and elution mobile phase
[0058] Liquid samples containing target proteins inevitably require the use of buffer solutions or mobile phases during loading, binding, and elution. The equilibrium buffer solution of the chromatography column, the sample loading buffer solution (the dialysate obtained by dialyzing the urine in the method of the present invention) and the mobile phase during elution may be the same or different. After optimization, the method of the present invention preferably uses the same equilibrium buffer solution, sample loading buffer solution and elution mobile phase. Because this does not cause changes in the chromatographic environment at the beginning of the chromatography, resulting in adverse effects on specific protein adsorption.
[0059] Equilibrium buffer solution, loading buffer solution, and elution mobile phase can be selec...
Embodiment 2
[0080] Example 2 Extraction methods of various urinary proteins
[0081] (1) Treatment of urine
[0082] Put 200ml of isolated urine from a healthy person into a semipermeable membrane, put it into a free diffusion dialysis device, soak it in pure water for dialysis overnight, and then put it into 50mM KH 2 PO 4 / K 2 HPO 4 The buffer solution (pH=6.5-7.2) was dialyzed overnight. Then, take 200ml of isolated urine from a healthy person after overnight dialysis and centrifuge it under a gravity of 10,000g-12,000g for 40min-60min to remove the sediment in the urine. Take the supernatant after centrifugation, and then use a 0.22 μm filter disc to install it on a syringe for filtration.
[0083] (2) Prepare the affinity chromatography column
[0084] 1. Preparation of Affinity Adsorbent
[0085] Wash Sepharose 4B (Sepharose 4B) with water, and disperse 20g of wet weight Sepharose 4B in 50ml of water, dissolve 200mg of Cibacron Blue F3-GA in 20ml of water, and shake gently ...
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