Purification method of hydrophobic polypeptide raw material

A purification method and hydrophobic technology, applied in the field of peptides, can solve the problems of low recovery rate and difficulty in dissolving hydrophobic peptides, and achieve the advantages of reducing solvent consumption, shortening operation time and operability, reducing energy consumption and production cost. Effect

Inactive Publication Date: 2018-02-16
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

[0005] In order to solve the above-mentioned technical problems, the purpose of the present invention is to provide a purification method that solves the problems of difficult dissolution and low recovery in the purification process of hydrophobic polypeptides

Method used

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  • Purification method of hydrophobic polypeptide raw material
  • Purification method of hydrophobic polypeptide raw material
  • Purification method of hydrophobic polypeptide raw material

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example 1

[0073] Dissolve 1 mg of Fmoc-cys(pal)-OH in 1 mL of a mixed solution of n-hexane and isopropanol (v:v=1:9), filter, and use a nitrile column Haobo CN 5 μm 20*250 mm for gradient detection analysis. The normal phase analysis conditions are: mobile phase A liquid is n-hexane, B liquid is isopropanol; time: 0-20min; flow rate: 1.00mL / min; A:B ratio: 99:1 to 5:95; wavelength: 220nm . Analysis results such as figure 1 As shown, it shows that the mobile phase is suitable for the analysis of Fmoc-cys(pal)-OH, and the peaks in the crude product can be well separated.

[0074] Dissolution and treatment of Fmoc-cys(pal)-OH crude product: 200mg of Fmoc-cys(pal)-OH crude product is ultrasonically dissolved with 20mL n-hexane-isopropanol mixed solution (v:v=1:9), after the dissolution is complete, centrifuge , and waited for sample preparation after filtration with a 0.45 μm organic phase filter membrane.

[0075] The preparative chromatographic column was LC3000I high performance liqui...

example 2

[0080] 1 mg of Fmoc-cys(pal)-OH was dissolved in 1 mL of n-hexane-isopropanol mixed solution (v:v=1:9), filtered, and gradient detection analysis was carried out using a nitrile column Haobo CN 5 μm 20*250 mm. The normal phase analysis conditions are: mobile phase A liquid is n-hexane, B liquid is isopropanol; time: 0-20min; flow rate: 1.00mL / min; A:B: 95:5 to 5:95; wavelength: 220nm . Analysis results such as Figure 5 As shown, it shows that the mobile phase is suitable for the analysis of Fmoc-cys(pal)-OH, and the peaks in the crude product can be well separated.

[0081] Dissolution and treatment of Fmoc-cys(pal)-OH crude product: 400mg Fmoc-cys(pal)-OH crude product was ultrasonically dissolved with 30mL n-hexane and isopropanol mixed solution (v:v=1:9), after the dissolution was complete, Centrifuge, and filter with a 0.45 μm organic phase filter and wait for sample preparation.

[0082] The preparative chromatographic column was LC3000I high performance liquid chroma...

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Abstract

The invention discloses a purification method of a hydrophobic polypeptide raw material. The purification method comprises steps of (1) polypeptide raw material dissolution, (2) normal-phase chromatographic analysis, and (3) chromatographic column preparation, adsorption, elution and collection. In step (1), a polypeptide raw material is dissolved into an organic solvent to obtain a solution of the polypeptide raw material. In step (2), the solution of the polypeptide raw material is connected to a normal-phase chromatographic column to carry out gradient detection to obtain analysis atlases.In step (3), according to the analysis atlases, the preparation gradient of chromatographic column is set, the solution of the polypeptide raw material is connected to the prepared chromatographic column to carry out adsorption, then the chromatographic column is eluted, and the eluent is collected to obtain purified hydrophobic polypeptide raw material. The purification method solves the problemsthat a large amount of a strongly hydrophobic raw material is hard to dissolve, the sample is not easy to elute in reverse phase preparation, and the recovery rate is low.

Description

technical field [0001] The invention relates to the technical field of polypeptides, in particular to a method for purifying raw materials of strongly hydrophobic polypeptides. Background technique [0002] Peptides are macromolecular substances with high activity, fast onset, high safety, and small side effects. Peptide drugs through biological expression or chemical synthesis play a huge role in the fields of anti-tumor, anti-virus, anti-bacterial, and vaccines. . With the development of medicine and biochemical technology, peptide drugs will become the most important drugs for monitoring, prevention, diagnosis and treatment in the 21st century. Its market share is constantly being eroded by biological products, among which peptide drugs are developing most rapidly. At present, nearly 60 peptide products have been approved for marketing in the world, and the global peptide drug market has exceeded 20 billion US dollars. Therefore, the study of peptide synthesis and puri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/16
CPCC07K1/16
Inventor 王锡平张中玉
Owner 苏州强耀生物科技有限公司
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