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Extraction and purification method of Gp96-polypeptide complex (CTL-OT)

A polypeptide complex and purification method technology, applied in the field of heat shock protein preparation, can solve the problems of sample size and quantity limitation, less Gp96-polypeptide complex, and limited clinical application, so as to improve content and efficiency, improve extraction efficiency, The effect of improving the purification efficiency

Pending Publication Date: 2020-12-22
和泓尚医(成都)生物科技有限公司
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Problems solved by technology

[0005] Although the clinical effect of using Gp96-polypeptide complex immunotherapy for various tumors is good, there are certain difficulties in obtaining Gp96-polypeptide complexes
So far, the source of Gp96-polypeptide complexes is mainly extracted and purified from tumor tissue resected from patients. Because clinical specimens of tumor tissue are not easy to obtain, sometimes even if tumor tissue is obtained, the sample size and quantity are limited. At the same time, Gp96 - The extraction and purification process of the polypeptide complex is relatively complicated. In many cases, the amount of Gp96-polypeptide complex extracted and purified is too small to achieve a sufficient amount for treating tumors. These conditions greatly limit its clinical application

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  • Extraction and purification method of Gp96-polypeptide complex (CTL-OT)

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Embodiment Construction

[0027] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. For the experimental methods without specific conditions indicated in the following examples, usually follow the conventional conditions or the conditions suggested by the manufacturer.

[0028] A method for extracting and purifying a Gp96-polypeptide complex, comprising the steps of:

[0029] 1. Pretreatment of isolated human placenta tissue:

[0030] (1-1) Thaw the cryopreserved isolated placenta tissue, remove the capsule material and connective tissue, then cut the placenta tissue into pieces, add an appropriate amount of pre-cooled pH=7.0, containing 30mM NaHCO 3 , 1mM lysate of phenylmethylsulfonyl fluoride, ground into a homogenate.

[0031] (1-2) Centrifuge the homogenate in a centrifuge at 10,000 rpm for 1 h at 4°...

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Abstract

The invention relates to the technical field of heat shock protein preparation, and particularly relates to an extraction and purification method of a Gp96-polypeptide complex. The method comprises the following steps: grinding human-derived in-vitro placenta tissue or tumor tissue into homogenate, then performing centrifugal extraction and agitation on the homogenate, and sequentially performingGp96 monoclonal antibody-Sepharose 4B affinity chromatography, HiTrap-Q Sepharose ion exchange chromatography extraction and purification on supernatant to obtain the Gp96-polypeptide complex. According to the method, the human-derived in-vitro placenta tissue or tumor tissue is used as a raw material, and the Gp96-polypeptide complex is obtained through the Gp96 monoclonal antibody-Sepharose 4B affinity chromatography and the HiTrap-Q Sepharose ion exchange chromatography, so that the extraction and purification content and efficiency of the Gp96-polypeptide complex are remarkably improved.

Description

technical field [0001] The invention relates to the technical field of heat shock protein preparation, in particular to a method for extracting and purifying a Gp96-polypeptide complex (CTL-OT). Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Cancer is a chronic disease that seriously endangers public health. There are more than 3.5 million new cancer cases and more than 2 million deaths in my country every year. Therefore, the situation of cancer prevention and control is very severe. The most common cancers in my country include lung cancer, breast cancer, gastric cancer, liver cancer, colorectal cancer, esophageal cancer, cervical cancer, and thyroid cancer. In recent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C07K1/18C07K1/22
CPCC07K14/47C07K2319/00
Inventor 王刚孟颂东吕祥涛
Owner 和泓尚医(成都)生物科技有限公司
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