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.

Pending Publication Date: 2020-05-01
瑞臻渼(北京)科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Currently, the urine proteins isolated and extracted from human urine are generally limited to four types: urokinase, human urinary trypsin inhibitor ulinastatin, albumin and β2-microglobulin, and a few other companies can extract them from the urine of pregnant women. Human chorionic gonadotropin (hCG for short) is isolated and extracted from human chorionic gonadotropin, but the raw material is not easy to obtain, so the production cost is relatively high
Currently known production techniques for separating and extracting urinary proteins from urine can only simultaneously extract at most two types of protein from a batch of urine, and generally can only separate and extract one type of urinary protein from the same batch of urine; and , these production processes are often crude and refined in one, and the purification of a urine protein can only be done from the beginning to the end, there is no room for choice, and the type and output of specific products produced cannot be effectively and flexibly adjusted according to market requirements

Method used

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  • Method for extracting various urinary proteins from urine
  • Method for extracting various urinary proteins from urine

Examples

Experimental program
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Effect test

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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Abstract

The invention provides a method for extracting various urinary proteins from urine. The method utilizes simultaneous affinity capture of various proteins in urine by a specific adsorbent, and staged specific elution from the adsorbent, and comprises the following steps: 1) collecting urine, and performing dialysis, centrifugation and filtration; 2) preparing a Cibacron Blue F3-GA-agarose gel affinity adsorbent, and performing column packing and equilibrating; 3) loading the urine and performing standing; and 4) performing elution in stages, and collecting elution fractions of each stage. The method of the invention can extract no less than eight kinds of high-value urinary proteins in parallel from the same batch of urine at one time, and significantly improves separation and extraction efficiency of urinary proteins, so that the urine is fully utilized as a specific biological resource.

Description

technical field [0001] The invention relates to protein extraction technology, more specifically, the invention relates to a method for extracting various urinary proteins from urine. Background technique [0002] A healthy adult excretes an average of 1.5 liters of urine per day, so the amount of urine excreted in a city every day (24 hours) is a very large number, and human urine contains as many as 1,500 kinds of proteins, many of which are It has quite high scientific research, medical and economic value. Therefore, if the urine protein can be separated, extracted and purified from human urine, the economic value will be considerable; moreover, the treated urine residue can also be used as an agricultural organic fertilizer, turning waste into treasure, and at the same time solving the problem of urine production. environmental problems. [0003] Currently, the urine proteins isolated and extracted from human urine are generally limited to four types: urokinase, human ...

Claims

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Application Information

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IPC IPC(8): C07K1/22C07K1/34
CPCC07K1/22C07K1/34
Inventor 陈彦涌方学敏
Owner 瑞臻渼(北京)科技发展有限公司
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