Apolipoprotein A1 purification method and ApoAI protein injection antigen

An apolipoprotein and purification method technology, applied in the field of biological separation, can solve the problems of limited sample volume, reduced detection accuracy, expensive processing equipment, etc., and achieves the effects of simple and novel method, low cost, and large amount of processed samples

Active Publication Date: 2015-04-15
深圳瑞亚力集团有限公司
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of this method, the processing equipment is expensive, the processing sample volume is limited, and the process time is long, resulting in the high cost of manufacturing ApoAI detection, and during the long-term processing, ApoAI is partly heterogeneous and degraded, which reduces the accuracy of detection.

Method used

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  • Apolipoprotein A1 purification method and ApoAI protein injection antigen
  • Apolipoprotein A1 purification method and ApoAI protein injection antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] In this example 1, the hydrophobic chromatography packing material phenyl sepharose high performance was used to treat the static human plasma.

[0050] S11, equilibrate the chromatographic column filled with phenyl sepharose high performance filler, with a height of 10cm and a diameter of 1.6cm, with equilibrium solution A (25mM Tris-HCl+25mM NaCl at pH 7.4), and equilibrate to the liquid outlet of the chromatographic column The pH value is the same as that of the balance solution;

[0051] S12, after the static human plasma is centrifuged at a low speed (4000g), the clear liquid after removing suspended particles is used as a sample for later use.

[0052] S21, after diluting 20ml of the sample in step S12 with plasma diluent (25mM Tris-HCl pH7.4) to 4 times the volume, directly load the sample on the above-mentioned chromatographic column equilibrated with equilibrium solution A. After the sample loading is completed, use the balance solution A to wash the unbound h...

Embodiment 2

[0060] S11, equilibrate the chromatographic column filled with phenyl sepharose high performance filler, with a height of 10cm and a diameter of 1.6cm, with equilibrium solution A (25mM Tris-HCl+25mM NaCl at pH 7.4), and equilibrate to the liquid outlet of the chromatographic column The pH value is the same as that of the balance solution;

[0061] S12, after the static human plasma is centrifuged at a low speed (4100g), the clear liquid after removing suspended particles is used as a sample for later use.

[0062] S21, after diluting 20ml of the sample in step S12 with plasma diluent (25mM Tris-HCl pH7.4) to 4 times the volume, directly load the sample on the above-mentioned chromatographic column equilibrated with equilibrium solution A. After the sample loading is completed, use the balance solution A to wash the unbound human plasma components in the chromatography filler; wherein, the control flow rate during the balance, sample loading and washing processes is 3ml / min; ...

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Abstract

The invention provides an apolipoprotein A1 purification method. The method is as below: obtaining a plasma sample, and clarifying the sample; conducting a first hydrophobic chromatography on the plasma sample after clarification, so as to obtain a first purification product; degreasing the first purification product; and conducting a second hydrophobic chromatography on the first purification product after degreasing treatment. The first purification elution includes twice elution, the first elution uses 5-10% organic alcohol, and the second elution uses 25-30% alcohol; and the second hydrophobic chromatography uses alcohol eluate with elution strength lower than 25-30%. The method of the invention uses strong hydrophobic mechanism of the lipid in an HDL structure for the first purification, and then the lipoprotein structure with destroyed HDL makes the apolipoprotein A1 separate from the lipid, so that hydrophobicity difference between the apolipoprotein A1 and complex protein with the same high hydrophobicity increases, hydrophobic separation occurs again, and apolipoprotein A1 with high purity can be obtained. The method is simple and novel, and has large sample treatment amount and high efficiency.

Description

technical field [0001] The invention belongs to the technical field of bioseparation, and in particular relates to a method for purifying apolipoprotein A1 and an ApoAI protein injection antigen. Background technique [0002] Apolipoprotein is a protein component that constitutes plasma lipoproteins, mainly divided into five categories: A, B, C, D, and E, and is an important component that constitutes plasma lipoproteins; its basic function is to form molecules with lipids and sterols Composite spheres to deliver lipids and sterols. Due to the above-mentioned basic characteristics and functions of apolipoprotein, it is often a structural and functional component of high-density lipoprotein (High Density Lipprotein, HDL). The formed high-density lipoprotein (High Density Lipprotein, HDL) is a kind of anti-atherosclerotic plasma lipoprotein, which is a protective factor for cardiovascular and cerebrovascular diseases. HDL is mainly synthesized in the liver (partially in the s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/775C07K1/20A61K38/17A61P9/10
CPCA61K38/00C07K14/775
Inventor 李光飞苏少博
Owner 深圳瑞亚力集团有限公司
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