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Method of preparing apolipoprotein A-I

An apolipoprotein and A-I technology, which is applied in the preparation methods of peptides, apolipoproteins, chemical instruments and methods, etc., can solve the problems such as being unsuitable for large-scale production of enterprises, high equipment requirements, and resource shortages, and achieve a comprehensive The effect of utilization, low equipment requirements, and blood resource saving

Inactive Publication Date: 2008-01-23
FUDAN UNIV
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Problems solved by technology

[0003] The disadvantages of this method are: 1. the raw material is human plasma, and there is a serious shortage of resources; 2. the production volume is small, and only about 25 mg of ApoA-I can be obtained each time the amount of separation (100mL plasma) is not suitable for large-scale production; Ultracentrifuge, etc.; ④ low yield, about 20%
[0004] So far, there is no method suitable for large-scale production of ApoA-I by enterprises

Method used

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  • Method of preparing apolipoprotein A-I

Examples

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Embodiment 1

[0048] 30g F-IV plus 120mL 65% ethanol-10mM NaHCO 3 (pH7.3) solution, stirred at -20°C for 2 hours, centrifuged to separate the supernatant, adjusted the pH of the supernatant to 5.5, centrifuged to separate the precipitate, and the precipitate was completely dissolved in 50mL 100mM Tris-HCl-6M Urea (pH8.6) solution. Add an equal volume of -20°C pre-cooled chloroform / ethanol (1:1) solution, shake evenly, and let stand at -20°C for 2h. Centrifuge, take the supernatant, add an equal volume of -20°C pre-cooled ethanol, shake evenly, let stand at -20°C for 2 hours, and centrifuge to separate the supernatant. The supernatant was concentrated and dialyzed against carbonate buffer (20 mM), pasteurized, sterilized and filtered, and freeze-dried to obtain ApoA-I dry powder.

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Abstract

The invention belongs to the pharmaceutical field of the biochemical drugs, which relates to the preparation method of the apolipoprotein A-I, in particular to a method for preparing the apolipoprotein A-I (Apo A-I) separated from the waste human plasma F-IV component. The invention adopts the waste human plasma F-IV component to gain the 98 per cent purity through the isoelectric point precipitate, chloroform and ethanol treatment and gain the 23.5 per cent yield based on the calculation of the content of the plasma F-IV Apo A-I, and keeps the apolipoprotein A-I of the natural Apo A-I biological activity; the wet weight / g can prepare 3.1mg purified Apo A-I. The invention lays good foundation for developing the Apo A-I medicinal value and provides a preparation method of human plasma F-IV produced industrially.

Description

technical field [0001] The invention belongs to the field of biochemical drug pharmacy, and relates to a preparation method of apolipoprotein A-I, in particular to a method for separating and preparing apolipoprotein A-I (ApoA-I) from discarded human plasma F-IV components. Background technique [0002] At present, there are only conventional laboratory preparation methods for the preparation of apolipoprotein A-I (ApoA-I). The method is to adjust the density of plasma to 1.34g / mL with KBr, add it to a centrifuge tube filled with liquid, and perform ultracentrifugation at 4°C, 166000×g, for 20 hours, and take the d1.063-1.021g / mL part (HDL), After dialysis and concentration, degrease with ethanol / ether, and centrifuge to separate the precipitate (HDL apolipoprotein). The supernatant is used to precipitate HDL apolipoprotein again with ether, and the two precipitations are combined. Bio-CAD rapid protein separator, POROS HQ -20 strong anion exchange resin separation to obtai...

Claims

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Application Information

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IPC IPC(8): C07K1/14C07K1/30C07K14/775
Inventor 吴满平廖雪玲
Owner FUDAN UNIV
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