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Preparation method of angiotensin converting enzyme inhibitor affinity column medium

A technology of angiotensin and converting enzyme, which is applied in the field of preparation of affinity column medium, and achieves the effects of stable purification method, wide source and easy access.

Inactive Publication Date: 2004-11-10
唐胜建
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Problems solved by technology

Since animals, plants and microorganisms in nature, especially Chinese herbal medicines contain a variety of antihypertensive substances including angiotensin-converting enzyme inhibitors, how to extract them quickly and effectively has become a bottleneck in the development of antihypertensive drugs. Therefore, In recent years, researchers at home and abroad have gradually increased the screening and purification of antihypertensive drugs for natural animals and plants. The purification method is usually to detect the inhibitory effect on angiotensin converting enzyme first, and then perform biochemical multi-step separation and purification. Thereby first prepare angiotensin-converting enzyme, "Biochemistry and Biophysics Progress" 2000; 27 (5) page 544-547 " separation and purification of porcine lung angiotensin-converting enzyme " article, open The preparation process of porcine lung angiotensin-converting enzyme by affinity chromatography gel is described. This paper only provides a method for the separation and purification of angiotensin-converting enzyme with a high yield. The affinity of angiotensin-converting enzyme inhibitors The preparation of column media has not been reported at home and abroad so far

Method used

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example 1

[0016] The preparation method of angiotensin-converting enzyme inhibitor affinity column medium comprises the following steps in sequence:

[0017] A. The preparation of the ligand. After cleaning the fresh pig lungs, remove the large blood vessels and fat, cut them into pieces, and add a 0.06mol / LTris-HCl buffer solution containing 0.25mol / L sucrose at a temperature of 4°C. The pH value of the solution was 7.2, and the weight-to-volume ratio of fresh pig lungs to the buffer was 3:10 in grams: milliliters, homogenized for 8 minutes, and centrifuged at 9000 × g for 30 minutes at a temperature of 4°C to obtain the supernatant, and then Carry out secondary precipitation of ammonium sulfate at a temperature of 0°C. The concentration of ammonium sulfate in the first stage precipitation is 1.6-2.0mol / L. After standing for 2 hours, centrifuge at 3000-4500×g for 1 hour to obtain the supernatant. The second stage The concentration of the precipitated ammonium sulfate is 2.1-2.6mol / L. A...

example 2

[0020] The preparation method of angiotensin-converting enzyme inhibitor affinity column medium comprises the following steps in sequence:

[0021] A. The preparation of the ligand. After cleaning the fresh pig lungs, remove the large blood vessels and fat, cut them into pieces, and add a 0.10mol / LTris-HCl buffer solution containing 0.25mol / L sucrose at a temperature of 4°C. The pH value of the solution was 7.6, and the weight-to-volume ratio of fresh pig lungs to the buffer was 3:15 in grams: milliliters, homogenized for 2 minutes, and centrifuged at 4000 × g for 45 minutes at a temperature of 4°C to obtain the supernatant, and then Carry out secondary precipitation of 1.6-2.6mol / L ammonium sulfate under the condition of 0°C at a temperature, the concentration of the first-stage precipitated ammonium sulfate is 1.6-2.0mol / L, centrifuge at 3000-4500×g for 1 hour after standing for 2 hours, and obtain The concentration of the supernatant and the second-stage precipitated ammoni...

example 3

[0024] The preparation method of angiotensin-converting enzyme inhibitor affinity column medium comprises the following steps in sequence:

[0025] A, the preparation of ligand, after cleaning the fresh pig lungs, remove the large blood vessels and fat, cut them into pieces, add the 0.08mol / LTris-HCl buffer solution containing 0.25mol / L sucrose at a temperature of 4°C, the buffer solution The pH value of the solution was 7.4, and the weight-to-volume ratio of fresh pig lungs to the buffer solution was 3:13 in grams: milliliters, homogenized for 5 minutes, and centrifuged at 7000 × g for 40 minutes at a temperature of 4°C to obtain the supernatant, and then Carry out secondary precipitation of 1.6-2.6mol / L ammonium sulfate under the condition of 0°C at a temperature, the concentration of the first-stage precipitated ammonium sulfate is 1.6-2.0mol / L, centrifuge at 3000-4500×g for 1 hour after standing for 2 hours, and obtain The concentration of the supernatant and the second-st...

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Abstract

An affinity column medium of angiotonin transferase inhibitor is prepared from fresh pig lung through homogenizing, centrifugal deposition to obtain supernatant, further classification deposition with ammonium sulfate, dialysis to obtain enzyme liquid, chromatography of enzyme peak part in gel column freeze-drying to obtain ligand, dissolving carrier and ligand in solution of sodium chloride, regulating pH value, adding cross-linking agent, shaking to fix ligand to carrier, and flushing with distilled water and sodium acetate. It can be used for purifying collagen and Chinese-medicine materials' active components.

Description

technical field [0001] The invention relates to a method for preparing an affinity column medium, more specifically to a method for preparing an angiotensin-converting enzyme inhibitor affinity column medium. Background technique [0002] Hypertension is one of the most important diseases that threaten people's lives at present. There are various kinds of drugs for treating hypertension, among which the drugs of angiotensin-converting enzyme inhibitors are more effective. Since animals, plants and microorganisms in nature, especially Chinese herbal medicines contain a variety of antihypertensive substances including angiotensin-converting enzyme inhibitors, how to extract them quickly and effectively has become a bottleneck in the development of antihypertensive drugs. Therefore, In recent years, researchers at home and abroad have gradually increased the screening and purification of antihypertensive drugs for natural animals and plants. The purification method is usually t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/00G01N30/00
Inventor 任维栋孙风祥唐胜建
Owner 唐胜建
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