A kind of preparation method of 1-1 type human plasma haptoglobin extract

A technology of binding haptoglobin and human plasma, applied in the field of protein separation and purification technology, it can solve the problems of protein structure, functional group damage, inability to extract haptoglobin, limited application scope, etc., to reduce the degradation of haptoglobin and increase the content , the effect of protecting structure and function

Active Publication Date: 2017-12-22
INST OF FIELD OPERATION SURGERY NO 3 MILITARY MEDICL UNIV PLA
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Problems solved by technology

The disadvantage of this method is that the cold ethanol precipitation method may cause damage to the protein structure and functional groups, and should be avoided as much as possible, and the above-mentioned polyethylene glycol 10000 crude extraction method is just based on the cold ethanol precipitation; in addition Before this method is extracted, there are very few types of miscellaneous proteins in the system, and it cannot be used to directly extract haptoglobin from plasma, so the scope of application is greatly limited

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  • A kind of preparation method of 1-1 type human plasma haptoglobin extract
  • A kind of preparation method of 1-1 type human plasma haptoglobin extract
  • A kind of preparation method of 1-1 type human plasma haptoglobin extract

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[0020] Human plasma comes from expired human plasma from blood banks, and is typed by SDS-PAGE electrophoresis or PCR typing, and other reagents are commercially available analytically pure products.

[0021] Prepare type 1-1 human plasma haptoglobin extract under 0°C ambient conditions:

[0022] 1) Place 50 ml of human plasma identified to contain haptoglobin type 1-1 in an ice bath, centrifuge at 8000 rpm for 10 minutes to remove condensed protein, and obtain 49 ml of supernatant;

[0023] 2) Add 11.074 grams of ammonium sulfate powder to the supernatant obtained in step 1, so that the final saturation of ammonium sulfate reaches 40% at 0° C., and centrifuge at 8000 rpm for 10 minutes after ice bathing for 20 to 30 minutes to obtain 46 milliliters of supernatant , add 7.038 grams of ammonium sulfate powder, so that the final saturation of ammonium sulfate reaches 65% at 0°C, and centrifuge at 8000 rpm for 20 minutes after ice bathing for 20 to 30 minutes, and take the precip...

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Abstract

The invention discloses a preparation method of a human plasma haptoglobin 1-1 extracting solution, which comprises the following step: sequentially carrying out ammonium sulfate fractional precipitation and polyethyleneglycol precipitation on haptoglobin 1-1-containing human plasma centrifugal supernatant preserved at low temperature so as to extract the human plasma haptoglobin 1-1 for fine purification. The purity of the human plasma haptoglobin 1-1 in the extracting solution prepared by the method is greater than 50%; and the method has the advantages of low-temperature operation and mild process, and is very suitable to be used as a crude extraction method for large-scale purification of haptoglobin 1-1 from human plasma.

Description

technical field [0001] The invention belongs to the technical field of protein separation and purification technology, and in particular relates to a preparation method of a 1-1 type human plasma haptoglobin extract in the protein separation and purification technology. Background technique [0002] Haptoglobin (Hp), also known as haptoglobin, is an acidic glycoprotein widely present in mammalian blood and other body fluids. It is mainly synthesized and degraded by the liver, and its content is regulated by lipopolysaccharide, cytokines, hormones, etc. An acute phase response protein. The haptoglobin molecule is composed of a and β subunits, and in most mammals (aβ) 2 There are three main subtypes of human haptoglobin, 1-1, 2-1, and 2-2, which are mainly caused by the diversity of a subunit. Human haptoglobin a subunit is encoded by alleles Hp1, Hp2, Hp1 encodes a 1 Subunit (molecular weight about 9.1kDa), Hp2 encodes a 2 subunit (molecular weight about 16kDa). Human ha...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C07K1/30
CPCC07K14/47
Inventor 刘良明臧家涛李涛
Owner INST OF FIELD OPERATION SURGERY NO 3 MILITARY MEDICL UNIV PLA
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