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Preparation method of specific endotoxin adsorption membrane and application thereof to field of assisted reproduction

A specific adsorption and endotoxin technology, applied in the field of assisted reproduction, can solve the problem of not being retrieved in the field of assisted reproduction, and achieve the effect of improving the embryo formation rate

Active Publication Date: 2018-09-28
成都艾伟孚生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

4. Promoting or inhibiting cell division, the difference in effect may be related to the source and concentration of endotoxin
There are relevant patents and literature reports on how to remove endotoxin in the pharmaceutical, equipment and other industries, but no relevant reports have been retrieved in the field of assisted reproduction

Method used

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  • Preparation method of specific endotoxin adsorption membrane and application thereof to field of assisted reproduction
  • Preparation method of specific endotoxin adsorption membrane and application thereof to field of assisted reproduction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Preparation of Endotoxin-Specific Adsorption Membrane

[0075] 1) Remove impurities

[0076] After the PVDF hollow fiber membrane (polyvinylidene fluoride membrane) was rinsed with 30% ethanol for 30 minutes, it was soaked in deionized water for 1 hour to wash away the impurities on the surface of the membrane.

[0077] 2) Hydrophilic modification

[0078] 4% (w / v) of KMnO 4 React with 3mol / l KOH mixture at 80°C for 30min with a constant flow peristaltic pump into the PVDF hollow fiber membrane module, and then use 5% (v / v) H 2 SO 4 and 5% (w / v) NaHSO 3 The mixture is restored.

[0079] 3) Coupled with HEC

[0080] After reduction treatment, the membrane was treated with 1mol / l NaOH and 1.5% (w / v) HEC mixture at 90°C for 10min to increase the hydrophilicity of the membrane surface. And put it in a constant temperature hot air drying oven at 90°C, and dry it with 0.5mol / l Na 2 CO 3 The solution was washed at 90°C for 10 min to wash off the unreacted HE...

Embodiment 2

[0088] Example 2 Sperm micromanipulation solution configuration

[0089] 1) The sperm micromanipulation base solution is filtered through the endotoxin-specific adsorption membrane, and repeated three times.

[0090] 2) Soak polyvinylpyrrolidone (PVP) with 0.75mol / L NaOH solution for 30min, then adjust the pH to neutral.

[0091]3) Steps 1) and 2) are mixed according to the ratio to obtain a low-endotoxin sperm micromanipulation solution.

Embodiment 3

[0092] Embodiment 3 effect detection

[0093] 1. Endotoxin detection method

[0094] The limulus reagent method was carried out in accordance with the "Pharmacopoeia of the People's Republic of China" 2015 edition "Bacterial endotoxin detection method (gel limit test)".

[0095] The adsorption capacity of the endotoxin-specific adsorption membrane of the present invention is tested.

[0096] The low-endotoxin sperm micromanipulation fluid of Example 2 was detected before and after using the endotoxin-specific adsorption membrane of the present invention.

[0097] 2. Test results

[0098] The endotoxin-specific adsorption membrane synthesized by the present invention has a maximum endotoxin adsorption capacity of 45ug / cm 2 .

[0099] The sperm micromanipulation fluid detection result of embodiment 2, see Table 1:

[0100] Table 1 Detection results of sperm micromanipulation fluid

[0101]

[0102]

[0103] Test results: before the treatment by the present invention,...

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Abstract

The invention belongs to the field of assisted reproduction and specifically relates to a specific endotoxin adsorption membrane. The specific endotoxin adsorption membrane disclosed by the inventionis specifically prepared by taking a PVDF hollow fibrous membrane as a carrier and polymyxin B as an adsorbent, wherein the polymyxin B is fixedly arranged on the carrier in a coupling way; the PVDF hollow fibrous membrane is coupled with HEC and is grafted with a hexamethylendiamine spacer arm; and the polymyxin B is coupled with the hexamethylendiamine spacer arm. The specific endotoxin adsorption membrane can be used for removing endotoxin in various buffer solutions, culture solutions, manipulation solutions and the like in the field of assisted reproduction and effectively reducing the content of endotoxin to an ideal range without affecting the content of effective components. The invention further provides an operating method for removing endotoxin in a sperm micromanipulation solution by using the membrane. The specific endotoxin adsorption membrane disclosed by the invention is used for removing endotoxin in the culture solutions in the field of assisted reproduction and is beneficial to assisted reproduction to increase the embryonic development rate and make an embryo form better.

Description

technical field [0001] The invention belongs to the field of assisted reproduction, and in particular relates to the preparation of an endotoxin-specific adsorption film coupled with polymyxin B and its application method in the field of assisted reproduction. Background technique [0002] Endotoxins are lipopolysaccharide substances (LPS) produced in the outer layer of the cell wall of Gram-negative bacteria, ie fragments of the bacterial corpse. Endotoxins generally exist in the form of polymers, and the chelation of phosphate groups with divalent metal ions can further stabilize the polymerization between endotoxin molecules and promote larger endotoxin complexes. The molecular weight of such complexes can range from several thousand to several Don't wait. Endotoxin has the characteristics of small size, strong thermal stability, and strong chemical stability, which make it difficult to be removed. It is now recognized that endotoxin is harmful to cell culture, and endo...

Claims

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Application Information

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IPC IPC(8): B01J20/26B01J20/28B01D71/34B01D69/08B01D69/02B01D67/00B01D15/08B01J20/30
CPCB01D15/08B01D67/0002B01D69/02B01D69/08B01D71/34B01D2325/12B01J20/24B01J20/265B01J20/28033
Inventor 余裕炉戴甄王瞿波夏晓伟米乐刘洪君严飞
Owner 成都艾伟孚生物科技有限公司
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