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Anti-sea snake venom serum nano-membrane filtering method

A filtration method, nano-membrane technology, applied in the preparation method of peptides, chemical instruments and methods, from serum immunoglobulin, etc., can solve the problems of high cost of nano-membranes, achieve accelerated industrialization, reduce costs, and increase protein content Effect

Pending Publication Date: 2021-12-17
SHANGHAI SERUM BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved in the present invention is to provide a nano-membrane filtration method for anti-sea snake venom. In order to solve the problem of high cost of the above-mentioned nano-membrane, through a large number of experiments, an optimal condition is obtained to improve the performance of the nano-membrane anti-venom serum process sample. Filtration flux, that is, increasing the total amount of protein that can be effectively filtered per unit membrane area, greatly reduces production costs, and is of great significance for improving the virus removal efficiency of anti-sea snake venom products and accelerating the application of nano-membrane filtration in the production of anti-sea snake venom

Method used

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  • Anti-sea snake venom serum nano-membrane filtering method

Examples

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Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 A nano-membrane filtration method for anti-sea snake venom

[0034] Proceed as follows:

[0035] One, according to the present invention figure 1 In the technical process, the preparation of anti-sea snake venom chromatography flow-through solution specifically includes the following steps;

[0036] 1. Obtain plasma;

[0037] 2. Pepsin digestion (pH3.5);

[0038] 3. The first precipitation of ammonium sulfate (the concentration of ammonium sulfate is 7% (mass volume ratio w / v));

[0039] 4. The second precipitation of ammonium sulfate (the concentration of ammonium sulfate is 30% (w / v));

[0040] 5. Plate and frame filter press;

[0041] 6. Alum adsorption (alum concentration is 6% (w / v));

[0042] 7. Plate and frame filter press;

[0043] 8. Concentrate and desalt the supernatant by ultrafiltration; in this ultrafiltration step, the ultrafiltration solution is 9.5mM phosphate buffer, and the pH value is 7.1;

[0044] 9. Ion-exchange column chromatogra...

Embodiment 2

[0050] Example 2 A nano-membrane filtration method for anti-sea snake venom

[0051] Proceed as follows:

[0052] One, according to the present invention figure 1 In the technical process, the preparation of anti-sea snake venom chromatography flow-through solution specifically includes the following steps;

[0053] 1. Obtain plasma;

[0054] 2. Pepsin digestion (pH4.0);

[0055] 3. The first precipitation of ammonium sulfate (the concentration of ammonium sulfate is 15% (w / v));

[0056] 4. The second precipitation of ammonium sulfate (the concentration of ammonium sulfate is 40% (w / v));

[0057] 5. Plate and frame filter press;

[0058] 6. Alum adsorption (alum concentration is 10% (w / v));

[0059] 7. Plate and frame filter press;

[0060] 8. Concentrate and desalt the supernatant by ultrafiltration; in this ultrafiltration step, the ultrafiltration solution is 9.5mM phosphate buffer, and the pH value is 7.1;

[0061] 9. Ion-exchange column chromatography; in this chr...

Embodiment 3

[0067] Example 3 A nano-membrane filtration method for anti-sea snake venom

[0068] Proceed as follows:

[0069] One, according to the present invention figure 1 In the technical process, the preparation of anti-sea snake venom chromatography flow-through solution specifically includes the following steps;

[0070] 1. Obtain plasma;

[0071] 2. Pepsin digestion (pH3.0);

[0072] 3. The first precipitation of ammonium sulfate (the concentration of ammonium sulfate is 3% (w / v));

[0073] 4. The second precipitation of ammonium sulfate (the concentration of ammonium sulfate is 20% (w / v));

[0074] 5. Plate and frame filter press;

[0075] 6. Alum adsorption (alum concentration is 0.2% (w / v));

[0076] 7. Plate and frame filter press;

[0077] 8. Concentrate and desalt the supernatant by ultrafiltration; in this ultrafiltration step, the ultrafiltration solution is 9.5mM phosphate buffer, and the pH value is 7.1;

[0078] 9. Ion-exchange column chromatography; in this chr...

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Abstract

The invention discloses an anti-sea snake venom serum nano-membrane filtering method which comprises the following steps: (1) preparing an anti-sea snake venom serum chromatography flow-through liquid, namely obtaining plasma; digesting with pepsase; precipitating ammonium sulfate for the first time; precipitating ammonium sulfate for the second time; performing plate-frame pressure filtration; alum adsorption; performing plate-frame pressure filtration; performing supernate ultrafiltration, concentration and desalination; performing ion exchange column chromatography; collecting chromatography flow-through liquid; (2) controlling the environment temperature of nano-membrane filtration to be 20-28 DEG C, and preparing the chromatography flow-through liquid into a solution with a certain concentration and a certain prescription; and (3) carrying out nano-membrane filtration according to the conditions in the step (2). The chromatography flow-through liquid in the anti-sea snake venom serum process is subjected to nano-membrane filtration virus removal, optimal conditions are obtained through multiple tests, the flux of nano-membrane filtration is increased, the production cost is greatly reduced, the virus removal efficiency of anti-sea snake venom serum products is improved, and the application of nano-membrane filtration in anti-sea snake venom serum production is accelerated.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and relates to a nano-membrane filtration method for anti-sea snake venom. Background technique [0002] Venomous snakebites are common in subtropical and tropical regions. In the rural areas of Asia, Africa, Latin America and Oceania, venomous snakebite is a particularly prominent public health problem, and the incidence of snakebite in my country shows a trend of increasing year by year. [0003] Sea snake venom has complex components and contains a variety of proteins, mainly including phospholipase A 2 , Three-finger toxin, metalloprotease, serine protease, vascular endothelial growth factor, snake venom factor, protease inhibitor, phospholipase B, etc. The components of sea snake venom are directly related to the clinical symptoms after sea snake bites, including muscle paralysis, respiratory paralysis, and muscle damage. [0004] The World Health Organization (WHO) emphasizes that antive...

Claims

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Application Information

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IPC IPC(8): C07K16/06C07K1/36C07K1/34C07K1/30C07K1/18
CPCC07K16/065C07K1/36C07K1/34C07K1/303C07K1/18
Inventor 陈则张黎明杨智柳国艳范铁炯王博何毅明王倩倩
Owner SHANGHAI SERUM BIOTECH
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