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Method for preparing endogenous digitalis factors

A digitalis, endogenous technology, applied to the field of preparation of endogenous digitalis factors, can solve the problems of difficulty in separation, too many impurities of similar molecular weight and the like

Active Publication Date: 2014-07-09
顺昊投资发展(天津)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention organically combines the ultrafiltration technology and the antibody technology, can specifically separate the endogenous digitalis-like factor from the tissue, and solves the problem of low content in the body and too many impurities of similar molecular weight for a long time. Difficult to separate

Method used

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  • Method for preparing endogenous digitalis factors
  • Method for preparing endogenous digitalis factors
  • Method for preparing endogenous digitalis factors

Examples

Experimental program
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Embodiment 1

[0030] Take blood samples from pregnant women, add an equal volume of methanol, mix well, centrifuge at 1500 rpm for 5 minutes, and let stand;

[0031] Take the supernatant, recover the methanol until it is exhausted, use an ultrafiltration membrane with a molecular weight cut-off of 10,000 to centrifuge for 10 minutes at 3,000 rpm, and retain the permeate;

[0032] Add an equal volume of digoxin antibody to the permeate, mix well, let stand at room temperature for 30 minutes, use an ultrafiltration membrane with a molecular weight cut-off of 10,000 for centrifugal ultrafiltration, and centrifugal ultrafiltration at 1000 rpm for 10 minutes, and keep the unpermeable Passed material, permeate discarded;

[0033] Add 0.3 mol / L glycine solution with volume equal to the volume of the non-permeated substance, mix evenly, centrifuge ultrafiltration with an ultrafiltration membrane with a molecular weight cut-off of 10,000, centrifuge at 1000 rpm for 10 minutes, pass through The solu...

Embodiment 2

[0047] (1) Take the placenta, cut it into pieces, freeze it with liquid nitrogen, and store it at -80°C until use;

[0048] (2) Put the slices into a homogenizer, put the homogenizer into the liquid nitrogen tank for 50 seconds, take out the homogenizer, and homogenize for 6 minutes at 1450 rpm;

[0049] (3) Take out the homogenate tissue, add an equal volume of methanol, mix well, centrifuge at 1450 rpm for 6 minutes, and let stand;

[0050] (4) Take the supernatant, recover the methanol until it is exhausted, use an ultrafiltration membrane with a molecular weight cut-off of 10,000 for centrifugal ultrafiltration, centrifuge at 2,800 rpm for 12 minutes, and retain the permeate;

[0051] (5) Add an equal volume of digoxin antibody to the permeate, mix well, let stand at room temperature for 25 minutes, use an ultrafiltration membrane with a molecular weight cut-off of 10,000 for centrifugal ultrafiltration, and perform centrifugal ultrafiltration at 800 rpm for 12 minutes. K...

Embodiment 3

[0054] (1) Take the placenta, cut it into pieces, freeze it with liquid nitrogen, and store it at -80°C until use;

[0055] (2) Put the slices into the homogenizer, put the homogenizer into the liquid nitrogen tank for 70 seconds, take out the homogenizer, and homogenize for 4 minutes at 1600 rpm;

[0056] (3) Take out the homogenized tissue, add an equal volume of methanol, mix well, centrifuge at 1600 rpm for 4 minutes, and let stand;

[0057] (4) Take the supernatant, recover the methanol to the end, use the ultrafiltration membrane with a molecular weight cut-off of 10,000 to centrifuge and ultrafilter, centrifuge at 3,000 rpm for 10 minutes, and retain the permeate;

[0058] (5) Add an equal volume of digoxin antibody to the permeate, mix well, let stand at room temperature for 35 minutes, use an ultrafiltration membrane with a molecular weight cut-off of 10,000 for centrifugal ultrafiltration, and perform centrifugal ultrafiltration at 1200 rpm for 8 minutes. Keep the n...

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Abstract

The invention belongs to the biotechnical field, and discloses a method for preparing endogenous digitalis factors. The method disclosed by the invention organically combines an ultrafiltration technology with an antibody technology to exclusively separate endogenous digitalis-like factors from tissues, so that the problem for a long time that the endogenous digitalis factors are hard to separate due to too low content of the endogenous digitalis factors in the tissues and too many impurities with similar molecular weights. The method disclosed by the invention has the characteristics of simple and convenient operational method, strong specificity and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of endogenous digitalis factor. Background technique [0002] Endogenous digitalis-like factor (EDLF), also known as endogenous digitalis-like factor or endogenous digoxin, is a biologically active substance produced and secreted in the body with digitalis activity. Pharmacological effects: [0003] (1) It has a strong heart effect. Studies have shown that EDLF can enhance the myocardial contractility of guinea pigs, and this effect is not affected by A or B adrenoceptor blockers. The level of EDLF in the plasma of patients with congestive heart failure is significantly increased, and there is a correlation with left atrial and pulmonary artery pressure. The content of total EDLF and free EDLF in pregnant women and fetuses is high, and twin pregnancy is significantly higher than that in singleton pregnancy. This may be due to the cardiotonic effect ...

Claims

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Application Information

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IPC IPC(8): A61K35/50
Inventor 李相国马洁
Owner 顺昊投资发展(天津)有限公司
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