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Method for purifying and preparing polypeptide

A separation and purification, acetonitrile technology, applied in the field of polypeptide purification and preparation, can solve the problems of volatile organic compound emission, difficult salt transfer, and high cost

Inactive Publication Date: 2016-04-20
上海吉尔多肽有限公司 +1
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  • Abstract
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Problems solved by technology

[0009] The purpose of the present invention is to provide a method for purifying and preparing polypeptides by using reversed-phase high-performance liquid chromatography combined with the recovery and recycling of acetonitrile, which mainly solves the problems of high cost, emission of volatile organic compounds, and salt conversion in existing polypeptide purification and preparation methods. Difficulties and other technical issues

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  • Method for purifying and preparing polypeptide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: refer to figure 1 ,

[0027] 1. Sample treatment: Weigh an appropriate amount of crude polypeptide and dissolve it in an aqueous solution of acetic acid and acetonitrile with a volume ratio: acetic acid: acetonitrile: water = 1:10:40, and perform ultrasonic treatment. After the sample is completely clarified, filter it with a filter membrane with a pore size of 0.45 μm. Collect the filtrate for later use.

[0028] 2. Separation and purification:

[0029] Purification conditions: Chromatographic column: CXTH-DAC200 (Beijing Innovation Tongheng Technology Co., Ltd. - dynamic axial compression column 200) with octadecylsilane bonded silica gel as the stationary phase, the column diameter and packing length are : 20cm×28cm. Mobile phase: A: 100% acetonitrile solution by mass percentage; B: 0.1% acetic acid aqueous solution by mass percentage. Flow rate: 1000ml / min. Detection wavelength: 230nm. Set the elution gradient and load the sample.

[0030] Purif...

Embodiment 2

[0033] Example 2: refer to figure 1 ,

[0034] 1. Sample treatment: Weigh an appropriate amount of crude polypeptide and dissolve it in an aqueous solution of acetic acid and acetonitrile with a volume ratio: acetic acid: acetonitrile: water = 1:10:40, and perform ultrasonic treatment. After the sample is completely clarified, filter it with a filter membrane with a pore size of 0.45 μm. Collect the filtrate for later use.

[0035] 2. Separation and purification:

[0036] Purification conditions: Chromatographic column: CXTH-DAC200 dynamic axial compression column with octadecylsilane bonded silica gel as the stationary phase, column diameter and packing length: 20cm×28cm. Mobile phase: A: 100% acetonitrile solution by mass percentage; B: 0.1% acetic acid aqueous solution by mass percentage. Flow rate: 1000ml / min. Detection wavelength: 230nm. Set the elution gradient and load the sample.

[0037] Purification process: Rinse the dynamic axial compression column with 10...

Embodiment 3

[0040] Example 3: refer to figure 1 ,

[0041] 1. Sample treatment: Weigh an appropriate amount of crude polypeptide and dissolve it in an aqueous solution of acetic acid and acetonitrile with a volume ratio: acetic acid: acetonitrile: water = 1:10:40, and perform ultrasonic treatment. After the sample is completely clarified, filter it with a filter membrane with a pore size of 0.45 μm. Collect the filtrate for later use.

[0042] 2. Separation and purification:

[0043] Purification conditions: Chromatographic column: CXTH-DAC200 dynamic axial compression column with octadecylsilane bonded silica gel as the stationary phase, column diameter and packing length: 20cm×28cm. Mobile phase: A: 100% acetonitrile solution by mass percent; B: 0.3% acetic acid aqueous solution by mass percent. Flow rate: 1000ml / min. Detection wavelength: 230nm. Set the elution gradient and load the sample.

[0044] Purification process: Rinse the dynamic axial compression column with 100% ace...

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Abstract

The invention relates to a method for purifying and preparing polypeptide. The technical problems that in an existing polypeptide purification and preparation method, the cost is high, and volatile organic compounds are discharged, and salt transfer is difficult are solved. According to the technical scheme, separation and purification, concentration, salt transfer and re-concentration are conducted on the polypeptide, produced high-concentration acetonitrile waste fluid is recycled by a system after distillation recovery is conducted, and the high-concentration acetonitrile waste fluid is synchronously applied to a polypeptide separation and purification method. The method for purifying and preparing the polypeptide greatly reduces the cost of polypeptide purification and preparation, and discharging of the acetonitrile waste fluid is reduced.

Description

technical field [0001] The invention relates to a method for purifying and preparing polypeptides, in particular to a method for purifying and preparing polypeptides by using reversed-phase high-performance liquid chromatography combined with recovery and recycling of acetonitrile. Background technique [0002] With the development of modern biotechnology, various peptide drugs have emerged, and peptides have great application value in clinical medicine. Since the invention of the solid-phase chemical synthesis of peptides, the chemical synthesis of various active peptides has made great progress, but the product components are relatively complex, generally consisting of several structurally similar peptide mixtures dominated by the target peptide. Therefore, the separation and purification of this mixture has always been an important research topic, and the optimization research on its separation and purification is particularly important. In addition, providing high-purit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/36C07K1/34C07K1/20C07K1/18
CPCC07K1/36C07K1/18C07K1/20C07K1/34
Inventor 张德志赵晶徐红岩
Owner 上海吉尔多肽有限公司
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