Method for preparing beta2 microglobulin crude product

A technology of β2-microglobulin and crude products is applied in the field of preparing β2-microglobulin, which can solve the problems of unsuitable industrial production, limited scale of preparation and high preparation cost, and achieve comprehensive utilization, reduce difficulty and prevent environmental problems. The effect of pollution

Active Publication Date: 2015-07-15
YANGZHOU AIDEA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The above studies are to a certain extent for the extraction and separation of β 2 -M provides certain methods, but although some of these methods can make the purity of the product reach a higher level, the available human urine is the urine of special populations, which is extremely difficult to collect, and the collection amount is very small. The scale is severely limited, not suitable for industrial production, and the preparation cost is relatively high

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  • Method for preparing beta2 microglobulin crude product
  • Method for preparing beta2 microglobulin crude product
  • Method for preparing beta2 microglobulin crude product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Take 100g UK crude product, add 1000ml of 0.01MTris-HCl buffer solution (pH7.6) to dissolve, centrifuge at 4000rpm for 10min, take the supernatant, filter it with a 0.45μm filter membrane, adjust the pH of the filtrate to 7.6, and conductance 1.06mS / cm, The well-balanced metal ion on the surface is Cu 2+ Metal chelate chromatography column (Chelating sepharose FF); and then the balance solution (0.3M NH 4 Cl, 0.01M Tris-HCl, pH 7.6) to wash the metal chelate chromatography column, collect the loading and washing breakthroughs for the preparation and purification of UK; use 0.07M glycine, 0.01MTris-HCl , pH8.0 to wash the column; use 0.5M glycine, 0.01M Tris-HCl, pH8.0 elution solution to elute the column; collect the elution solution.

[0035] Add ammonium sulfate powder to the collected eluent to saturation, stir while adding, let it stand for 4 hours, add 10 g of diatoms to collect by centrifugation and dry the precipitate to obtain β 2 -M crude product 16 g.

[00...

Embodiment 2

[0040] Take 100g of crude UTI, add 1000ml of 0.1M phosphate buffer (pH6.0) to dissolve, filter through diatomaceous earth plate and frame, and then filter with 0.45μm filter membrane. Adjust the pH of the filtrate to 6.0, the conductance is 0.2mS / cm, and the balanced metal ion is Zn 2+ The metal chelation chromatography column (Chelating sepharose FF); then use the equilibrium solution (equilibrium solution formula: 0.1MNH 4 Cl, 0.1M phosphate, pH6.0) to wash the metal chelate chromatography column, collect the sample breakthrough and wash the breakthrough for the preparation and purification of UTI; use 0.05M glycine, 0.1M phosphate, Wash the column with pH 7.0 flushing solution; elute the column with an eluting solution containing 0.1M glycine, 0.02M Tris-HCl, pH 7.0; collect the eluting solution.

[0041] Add ammonium sulfate powder to the collected eluted solution to saturation, stir while adding, let it stand for 4 hours, add 10 g of diatomaceous earth to collect by cent...

Embodiment 3

[0046] Take 100g of crude HMG, add 1000ml of 0.05M phosphate buffer (pH7.0) to dissolve, centrifuge at 4000rpm for 10min, take the supernatant, and filter it with a 0.45μm filter membrane to obtain the filtrate. Adjust the pH of the filtrate to 7.0, the conductance is 0.78mS / cm, and the balanced metal ion is Cu 2+ The metal chelate chromatography column (Chelating sepharose FF); then use the equilibrium solution (0.2MNH 4 Cl, 0.05M Phosphate, pH7.0) to wash the metal chelate chromatography column, collect the sample penetration and washing breakthrough, for the preparation and purification of HMG; use 0.1M glycine, 0.05M phosphate, Wash the column at pH 7.6; elute the column with an elution solution containing 0.38M glycine, 0.05M Tris-HCl, pH 8.3; collect the elution solution.

[0047] Add ammonium sulfate powder to the collected eluted solution to saturation, stir while adding, let it stand for 4 hours, add 10 g of diatomaceous earth to collect by centrifugation and dry the...

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Abstract

The invention discloses a method for preparing a beta2 microglobulin crude product. The method comprises the following steps: dissolving a UK crude product, a UTI crude product, an HMG crude product or an HCG crude product; centrifuging and filtering by a sheet frame; adding to a metal chelating chromatography column which is balanced by balance liquid, wherein the UK, the UTI, HMG and HCG are not adsorbed but beta2-M is combined to the metal chelating chromatography column, so as to realize separation; collecting a beta2 microglobulin elution peak, and preparing a beta2 microglobulin crude product; collecting and sampling, washing and penetrating, and finally preparing and purifying UK, UTI, KMG or HCG. By adopting the method, the beta2 microglobulin can be extracted from normal human urine in a large scale by coproduction of the beta2 microglobulin and these urine proteins. Thus, comprehensive utilization of the urine is realized, and the production cost of the beta2 microglobulin is greatly reduced.

Description

Technical field [0001] The invention involves the purification method of protein, and it is more specifically to prepare β from the urine of normal people 2 -The method of microfin protein. Background technique [0002] β 2 Microquarin (β 2 -Microglobulin, referred to as β for short 2 -M) It was the first time from the Swedish scientist Berggard in 1968 from the separation of the urine, the molecular weight was 11800, and the electric point was 5.7. 2 The regional name is named.β 2 -M is a single-chain polypeptide composed of 99 AA. It exists widely in plasma, urine, cerebral spinal cord, saliva and colostrum.Essenceβ 2 -M is rejuvenated in the renal tube, but when the renal tube is damaged, the concentration in the urine will increase significantly, and the β in the blood or urine is detected in clinical practice. 2 -M concentration is the clinical diagnosis of clinical renal function measurement, kidney transplantation, diabetic nephropathy, heavy metal cadmium, mercury poisoni...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C07K1/16C07K1/14
CPCC07K14/47
Inventor 苗丕渠居维艳
Owner YANGZHOU AIDEA BIOTECH
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