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Method for extracting and separating IgM and IgG from plasma

A technology for plasma and gel-free plasma, applied in the field of extraction and separation of IgM and IgG, can solve the problems of not separating IgM protein separately, low IgG efficiency and high production cost, and achieves improved extraction and separation efficiency, reduced production cost, and reduced production costs. energy consumption effect

Active Publication Date: 2020-11-20
HUALAN BIOLOGICAL ENG CHONGQING +2
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  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the deficiencies in the prior art, the present invention provides a method for extracting and separating IgM and IgG from plasma to solve the problem that the low-temperature ethanol precipitation method for separating IgG has low efficiency and does not separate IgM protein. Usually, it needs to be carried out at minus 20°C, which consumes more energy, and ethanol is a flammable and explosive substance. For safety reasons, explosion-proof equipment must also be installed in the workshop, and the production cost is relatively high.

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  • Method for extracting and separating IgM and IgG from plasma
  • Method for extracting and separating IgM and IgG from plasma
  • Method for extracting and separating IgM and IgG from plasma

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Embodiment Construction

[0015] The present invention will be described in further detail below in combination with the accompanying drawings, and specific implementation methods will be given.

[0016] A method for extracting and separating IgM and IgG from plasma. It is characterized in that, comprising the following steps:

[0017] S1. Plasma preparation

[0018] Melt the fresh frozen plasma, centrifuge continuously at 0-3°C to remove the cold glue, and the obtained supernatant is the cold glue-free plasma, and then use a filter element with a pore size of 0.22 μm to filter the cold glue-free plasma. Clarified cold-depleted plasma was obtained. Most of the coagulation factors and lipoproteins are removed by centrifugal precipitation in a low temperature environment, so that the subsequent steps can better extract the required immunoglobulins.

[0019] S2. Preparation for anion exchange chromatography

[0020] The chromatographic filler of anion exchange chromatography is a strong anionic filler...

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Abstract

The invention belongs to the field of protein extraction, and specifically discloses a method for extracting and separating IgM and IgG from plasma. The pH value of cold-glue removed plasma is adjusted to 6.3-7.0, and the conductivity of cold-glue removed plasma is adjusted to 9-12 mS / cm, anion exchange chromatography is carried out to obtain a first flow-through liquid and an anionic gel packingwith IgM protein adsorbed; and the pH value of the first flow-through liquid is adjusted to 5.4-5.6, the conductivity of the first flow-through fluid is adjusted to 2.0-4.0 mS / cm, and cation exchangechromatography is performed to obtain a second flow-through fluid and a cation gel packing with IgG protein adsorbed. By adopting the scheme of the invention, the problem of lower separation efficiency of IgM and IgG can be solved.

Description

technical field [0001] The invention belongs to the field of protein extraction, and in particular relates to a method for extracting and separating IgM and IgG from blood plasma. Background technique [0002] Immunoglobulin is a glycoprotein molecule produced by plasma cells stimulated by antigens under various physiological and pathological conditions. It usually has a Y-shaped structure formed by two heavy chains and two light chains. Immunoglobulins can be divided into IgM, There are five types of IgG, IgD, IgA, and IgE, among which the content of IgG in plasma is 7-16g / L, accounting for 75%-80% of the total immunoglobulin; IgM is extremely strong in the antibody-mediated humoral immune response It is the first antibody produced by the body after anti-infection and immune stimulation. [0003] For the outbreak of novel coronavirus pneumonia since December 2019, the diagnosis and treatment plan issued by the National Health Commission mentions that the diagnostic criteri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/06C07K1/18C07K1/14
CPCC07K16/065
Inventor 肖岚刘余江张海梦任虹滕世超张宝献
Owner HUALAN BIOLOGICAL ENG CHONGQING
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