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Purification method of RGD peptide containing multiple cysteines

A purification method, cysteine ​​technology, applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve problems such as easily oxidized RGD sequence polypeptides, shorten operation time and operability, reduce production costs, and increase yield Effect

Active Publication Date: 2020-01-07
苏州强耀生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to solve the above-mentioned technical problems, the purpose of the present invention is to provide a method that overcomes the deficiencies in the existing production process and solves the problem that the RGD polypeptide containing multiple thiols is easy to purify during the purification process. Purification method of RGD sequence polypeptide with oxidation problem

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  • Purification method of RGD peptide containing multiple cysteines
  • Purification method of RGD peptide containing multiple cysteines
  • Purification method of RGD peptide containing multiple cysteines

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Embodiment Construction

[0064] The present invention will be further described below in conjunction with the accompanying drawings.

[0065] The following examples are further detailed descriptions of specific embodiments of the present invention. Not all examples. Other embodiments based on the present invention made by professionals in the field without creative work all belong to the protection scope of the present invention. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0066] RGD sequence polypeptide SEQ ID No.1: CCRGDKGPDC (Disulfide Bridge: C2-C10)

[0067] L-cysteinyl-L-cysteinyl-L-arginyl-L-glycyl-L-aspartyl-L-lysyl-L-glycyl-L-prolyl -L-Aspartyl-L-cysteine ​​(cysteinyl at position 2 and cysteine ​​sulfhydryl group at position 10 form an intramolecular disulfide bond)

[0068] Molecular weight: 1051.19

[0069] Molecular formula: C38H62N14O15S3

[0070] Structure diagram:

[0071]

[0072] 1 m...

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Abstract

The invention provides a method for purifying an RGD peptide containing a plurality of cysteines, the RGD cyclic peptide is characterized by containing two or more cysteines, an -RGD-sequence, and various modified peptides on such basis. A dissolving step comprises: dissolving polypeptide in a mixed solvent of water and acetonitrile to obtain a polypeptide solution, and adding a small amount of reducing reagent into the polypeptide solution. A reversed-phase chromatographic column analysis step comprises connecting an RGD polypeptide solution to a reversed-phase chromatographic analysis columnfor gradient detection to obtain an analysis spectrum. Reversed-phase preparative chromatographic column adsorption and elution collection comprises setting the preparation gradient of the preparative chromatographic column according to the analysis spectrum, connecting the RGD polypeptide solution to the preparative chromatographic column for sample injection adsorption and elution collection, and implementing elution collection. A preparation mobile phase needs to be subjected to degassing and nitrogen filling. The problem that the RGD peptide tends to oxidation and isomer formation after oxidation in the purification process is solved, and products with the purity of more than 98% can be obtained.

Description

technical field [0001] The invention relates to the technical field of polypeptides, in particular to a purification method for -RGD-polypeptides containing multiple cysteines which are easily oxidized. Background technique [0002] The incidence of cancer in the world has shown a sharp upward trend in recent years. Studies have found that the growth of tumors depends on the formation of blood vessels. Small peptides containing RGD sequences are membrane-penetrating peptides, which can cross the cell membrane and inhibit angiogenesis, thereby inhibiting the formation, growth and metastasis of tumor cell blood vessels. . Therefore, the RGD polypeptide plays an important role in cancer treatment and has great market value. At present, the research on peptides containing RGD sequences is becoming more and more mature, and some derivative peptides of RGD peptides have appeared, such as paclitaxel, adamantanecarboxylic acid, gemcitabine, etc. coupled to RGD peptides, and there ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/14G01N30/34G01N30/89
CPCG01N30/06G01N30/14G01N30/34G01N30/89G01N2030/146Y02P20/55
Inventor 王锡平
Owner 苏州强耀生物科技有限公司
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