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Separation and purification method of multi-part mixed human plasma IgG sample simulating IVIg

A separation and purification technology with multiple servings, applied in the field of biomedicine, can solve difficult IVIg and other problems, and achieve high sample uniformity, high repeatability, and good enrichment effect

Active Publication Date: 2021-03-30
BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many defects and shortcomings of IVIg have been limited by industrialization for a long time, making it difficult to carry out relevant research on IVIg editing optimization required for basic research on products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] A method for separating and purifying a multi-part mixed human plasma IgG sample simulating IVIg, which comprises the following steps of mixing several parts of plasma, mixing 50% saturated ammonium sulfate method for crude purification, crude protein ultrafiltration, and crude protein ion exchange chromatography. step, purified protein ultrafiltration step, purified protein buffer system replacement and component detection step, to obtain multi-dose mixed high-purity and high-abundance IgG samples simulating IVIg products for use in downstream research.

[0038] Specific steps are as follows:

[0039]Step 1: Mix several parts of plasma;

[0040] Step 2: Coarse purification by mixing 50% saturated ammonium sulfate method:

[0041] (1) The mixed slurry obtained in step 1 and the saturated ammonium sulfate solution with a saturation of 100% are mixed in equal volumes at a volume ratio of 1:1 under normal temperature conditions, centrifuged after standing, and the superna...

Embodiment 2

[0055] A method for separating and purifying a multi-part mixed human plasma IgG sample simulating IVIg, the method comprises the following steps: a pretreatment step of mixing several parts of plasma, a slurry mixing step for removing cryoprecipitation, a slurry mixing 50% saturated ammonium sulfate method for crude purification, and a crude protein ultra-precipitation step. Filtration step, crude protein ion exchange chromatography purification step, purified protein ultrafiltration step, buffer system replacement of purified protein and component detection steps, to obtain a high-purity and high-abundance IgG sample of multiple slurries simulating IVIg products for downstream use Research use.

[0056] Specific steps are as follows:

[0057] Step 1: Pretreatment of multi-part and human-mixed plasma: to obtain raw plasma, according to research needs, any number of plasma parts, type of plasma source, mixing ratio, type and quantity of additives can be used to prepare mixed p...

Embodiment 3

[0074] Other contents are as in Example 2, wherein the elution buffer (Elution Buffer; EB) is a 0.22 μm filter membrane ultrafiltration phosphate buffer (PB) containing 0.2 M arginine, pH=7.5. The precipitation was mixed with a volume ratio of 1:1 to redissolved crude protein, and an appropriate amount of equilibration buffer (Start Buffer; SB) was added to the precipitation, followed by shaking and mixing to fully dissolve the precipitate.

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PUM

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Abstract

The invention belongs to the field of biomedical technology research and development, and relates to a separation and purification method of a multi-part mixed human plasma IgG sample simulating IVIg.The method includes the steps of a plurality parts of plasma are mixed, and the mixed plasma is successively subjected to treatments including 50% saturated ammonium sulfate method crude purification, crude protein ultrafiltration, crude protein ion exchange chromatography purification, purified protein ultrafiltration, purified protein buffer system replacement and component detection to obtainthe high-purity and high-abundance multi-part mixed human plasma IgG sample simulating IVIg, which can be used in following steps. The IgG content of the sample is high, the IgG spectrum is wide, thehydrophilicity and the body affinity are high, and the method is specially used for research requirements of product improvement and the like of plasma purified protein products such as IVIg and the like.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and particularly relates to a method for separating and purifying a multi-part mixed human plasma IgG sample simulating IVIg. Background technique [0002] Intravenous human immunoglobulin pH4 (intravenous immunoglobulins, IVIG) is an important human plasma protein product whose main component is immunoglobulin IgG. [0003] As a traditional blood product, IVIg has been widely used because of its safe and effective characteristics, and has become one of the traditional blood products with a high degree of industrialization. But it is also limited by its high degree of industrialization, and the editability of commercial IVIg products is extremely low. IVIg has a number of defects and shortcomings for a long time. Due to the limitation of industrialization, it is difficult to carry out relevant research on IVIg editing and optimization required for basic research on products. [0004] How to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/06C07K1/36C07K1/34C07K1/30C07K1/22C07K1/18
CPCC07K16/065Y02A50/30
Inventor 马莉徐继轩张巍杜晞王宗奎叶生亮张容王娅李长清
Owner BLOOD TRASFUSION INST CHINESE ACAD OF MEDICAL SCI
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