Method for separating and purifying pig plasma globulin

A technology for separation and purification of globulin, applied in plasma globulin/lactoglobulin, peptide preparation methods, chemical instruments and methods, etc., can solve the problems of large-scale preparation that have not yet been seen, and avoid the impact on product quality and the environment The effect of destruction, increasing product added value, and high separation efficiency

Inactive Publication Date: 2019-11-19
HENGYANG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Porcine plasma globulin has multiple uses, and there is no large-scale preparation technology for it

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A method for separating and purifying porcine plasma globulin, the method comprising the following steps:

[0037] Step 1. Preparation of globulin-rich precipitate:

[0038] a. Heregulin content: Measure the protein content in 1L plasma by biuret method, dilute the plasma with 0.9% sodium chloride until the protein content is 5%;

[0039] b. Adjust pH value and temperature: Detect the pH value of the diluted plasma, adjust the pH to 6.5 with 1M / L citric acid, adjust the temperature to 1°C, and add ethanol;

[0040] c. After adding ethanol, adjust the pH to 6.6;

[0041] d. Stirring reaction: the temperature is controlled at -4.5°C, and the reaction is stirred for 2 hours;

[0042] e. Rinse the centrifuge cup with 14.5% ethanol balance solution, pre-cool the centrifuge cup to -3.0°C, centrifuge the product, take the supernatant, cool the supernatant to -4.8°C, filter, and set aside;

[0043] Step 2, the separation of globulin:

[0044] a. Production and separation of...

Embodiment 2

[0058] Embodiment two: a kind of method for separating and purifying porcine plasma globulin, this method comprises the following steps:

[0059] Step 1. Preparation of globulin-rich precipitate:

[0060] a. Heregulin content: Measure the protein content in 1000mL plasma by biuret method, dilute the plasma with 0.9% sodium chloride until the protein content is 5%;

[0061] b. Adjust pH value and temperature: Detect the pH value of the diluted plasma, adjust the pH to 6.6 with 1M / L citric acid, adjust the temperature to 1.5°C, and add ethanol;

[0062] c. After adding ethanol, adjust the pH to 6.7;

[0063] d. Stirring reaction: the temperature is controlled at -4.6°C, and the stirring reaction is carried out for 2 hours.

[0064] e. Rinse the centrifuge cup with 14.5% ethanol balance solution, pre-cool the centrifuge cup to -4.0°C, centrifuge the product, take the supernatant, cool the supernatant to -5.0°C, filter, and set aside;

[0065] Step 2, the separation of globulin...

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Abstract

The invention discloses a method for separating and purifying pig plasma globulin, wherein the method comprises the steps: step 1, preparation of a precipitate rich in globulin: adjusting the proteincontent to 5%; adjusting the pH value and temperature; adjusting the pH after completion of addition of ethanol; carrying out stirring reaction; and swing to wash a centrifugal cup with a 14.5% ethanol balance solution, precooling the centrifugal cup, centrifuging the product, taking a supernatant, and cooling the supernatant to -4.8 to -5.5 DEG C, and filtering for standby application; step 2, separation of the globulin: preparing and separating the globulin; and carrying out press filtration separation of the globulin; and step 3, purification of the globulin. The extraction process is established for the pig plasma globulin, and the obtained product is a single protein; by adjustment and control of the temperature, pH and other parameters, and comprehensive use of the ethanol precipitation and dialysis technology, the target protein has high purity and high extraction rate; the price of the used materials is low, so the cost is greatly reduced; the whole separation and purificationprocess takes less than 10 h, so the separation efficiency is high; the pig plasma globulin is a high value added product and the utilization value of pig blood can be significantly improved.

Description

technical field [0001] The invention relates to the technical field of globulin separation and purification, in particular to a method for separating and purifying porcine plasma globulin. Background technique [0002] Globulin is an immune protein widely present in human or animal blood, also known as immunoglobulin. When the immune system encounters a foreign invader, it will produce different amounts of globulin according to the different invaders. If the invader is difficult to eliminate, the immune system will stimulate the lymph to produce more globulin until the invader is eliminated. . [0003] At present, human globulin mainly comes from the venous blood of healthy people, among which there are several kinds of gamma globulin preparations according to the protein content of 10%, 16%, 16.5% (the concentration of domestic products is above 10%), of which gamma globulin accounts for 95% The other is placental blood-derived gamma globulin (human placental blood gamma ...

Claims

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Application Information

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IPC IPC(8): C07K14/47C07K1/14C07K1/36C07K1/30C07K1/16C07K1/18
CPCC07K14/4717
Inventor 杨海唐青海王芳宇杨灿刘会敬唐姣玉龚慧芳张淇美
Owner HENGYANG NORMAL UNIV
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