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Polypeptide and preparation method thereof

A host cell and nucleic acid sequence technology, applied in the field of polypeptides and their preparation, can solve the problems of low hemolysis rate, unsatisfactory antibacterial activity and stability of natural polypeptides, and high toxicity, and achieve low hemolysis, strong broad-spectrum antibacterial activity, The effect of low production cost

Pending Publication Date: 2022-02-11
杭州长龄生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problems of unsatisfactory antibacterial activity and stability of existing natural polypeptides, and high toxicity, the present invention provides a series of polypeptides with high biological activity, low hemolysis rat

Method used

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  • Polypeptide and preparation method thereof
  • Polypeptide and preparation method thereof
  • Polypeptide and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0070] The design, synthesis and purity detection of embodiment 1 polypeptide SEQ ID NO.1-5

[0071] According to the amino acid sequence of the mature peptide of the natural polypeptide BAMP 28 (No. OCB93651.1), the amino acid sequence of the polypeptide SEQ ID NO. 1-5 of the present invention is artificially designed, and the polypeptide is synthesized by chemical solid-phase synthesis.

[0072] Adopt HPLC to detect the synthetic polypeptide purity, confirm that the synthetic polypeptide purity is all greater than 95%; HPLC detection condition: mobile phase A is to contain the trifluoroacetic acid of 0.05% (V / V); Mobile phase B is to contain 0.05% (V / V) ) of acetonitrile, the detection wavelength is 210nm, and within 40 minutes, the mobile phase B increases linearly from 0 to 90% (V / V).

Embodiment 2

[0074] The difference between this example and Example 1 is that the amino acid sequence SEQ ID NO.6 of the polypeptide SEQ ID NO.6 is artificially designed. All the other are identical with embodiment 1.

Embodiment 3

[0076] The difference between this example and Example 1 is that the amino acid sequence SEQ ID NO.7 of the polypeptide SEQ ID NO.7 is artificially designed. All the other are identical with embodiment 1.

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Abstract

The invention provides a polypeptide and a preparation method thereof, and relates to the technical field of biological pharmacy. Artificial design and modification are carried out on the basis of polypeptide from a natural source, the polypeptide shown in the amino acid sequence of SEQ ID NO.1-5 is provided, and the blood solubility of the polypeptide is reduced on the basis of keeping the biological activity of the polypeptide. The preparation method comprises the following steps: S1, artificially designing an amino acid sequence of the polypeptide and a nucleotide sequence for coding the amino acid sequence; S2, connecting the nucleotide sequence with an expression vector, transforming and/or transfecting into a host cell, and screening positive clones; S3, culturing positive clones, and inducing polypeptide expression; and S4, breaking the host cell, and purifying the protein polypeptide to obtain the polypeptide. The preparation method is high in expression efficiency, simple in separation and purification process and high in product purity, the purity of the obtained polypeptide can reach 98%, the pharmaceutical industrialization level is reached, the production cost is low, and the preparation method is suitable for large-scale industrial production.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and specifically relates to a polypeptide and a preparation method thereof. Background technique [0002] With the widespread use of antibiotics, the phenomenon of abuse has generally increased, accompanied by the continuous increase of drug-resistant strains, making anti-infection treatment fall into a crisis of drug resistance. When synthetic quinolone antibacterial drugs came into the market, clinical pathogenic bacteria were very sensitive to this new type of antibacterial drug, and there were almost no drug-resistant strains. After years of extensive use, the resistance rate of clinical pathogenic bacteria to this type of synthetic antibacterial drug increased rapidly. For example, the resistance of Escherichia coli to quinolones is as high as 70%. In order to deal with drug-resistant bacterial infections, the existing technology is on the one hand structurally modifying traditio...

Claims

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Application Information

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IPC IPC(8): C07K7/08C12N15/11C12N15/70C12N1/21C12Q1/689C12Q1/6895G01N33/569A61K38/10A61P31/04A61P31/10C12R1/19
CPCC07K7/08C12N15/70C12Q1/689C12Q1/6895G01N33/569A61K38/10A61P31/04A61P31/10
Inventor 史爱连杨冬林坤晓
Owner 杭州长龄生物科技有限公司
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