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Polypeptide for prolonging blood circulation time of bacteriophage

A technology of blood circulation time and bacteriophage, which is applied in the field of biomedicine to achieve the effect of prolonging blood circulation time

Active Publication Date: 2018-10-26
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nevertheless, the above scheme still has its limitations, and the application of more innovative strategies to obtain long-term circulating phages with better antibacterial effects remains to be broken through.

Method used

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  • Polypeptide for prolonging blood circulation time of bacteriophage
  • Polypeptide for prolonging blood circulation time of bacteriophage
  • Polypeptide for prolonging blood circulation time of bacteriophage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Screening of specific time-delayed short peptides

[0054] In this example, the library used to screen specific time-delayed short peptides is provided by New England Biolabs, which is a heptapeptide library containing disulfide bonds (Ph.D. TM -C7C). Random fragments in this library are flanked by cysteine ​​residues that can be oxidized during phage assembly to form disulfide linkages, thus forming a cyclic peptide that interacts with the target. This library contains more than two billion clones. Random peptides in the library are amino-terminal to the small coat protein pIII, so five copies are expressed per phage particle. Ph.D. TM The position of the phage-expressed random sequence in the -C7C library is preceded by alanine-cysteine. There is a short linker sequence (glycine-glycine-glycine-serine) between the random peptide and the pIII protein. (Ph.D. TM - C7C Phage Peptide Display Kit, http: / / www.neb.com / nebecomm / products / productE8120.asp).

...

Embodiment 2

[0061] Example 2: Delay Capability Test

[0062] Take 1×10 respectively 11 A phage carrying the displayed peptides BCP1, BCP2, and BCP6 obtained in Example 1 was mixed with random phage SC (displayed peptide sequence: CNATLPHQC, SEQ ID NO: 4), and injected into the rat body through the tail vein, respectively, at 0h (3min). ) and 72h to take blood plate to detect phage.

[0063] figure 2 The number of phages at 0h and 72h are shown. It can be seen from the figure that the numbers of the four phages are similar at 0 h, but the SC phages have disappeared at 72 h, and the proportion of BCP1 is the highest, followed by BCP6 and BCP2. It can be seen that the phages carrying the three displayed peptides obtained in Example 1 all have the ability to significantly prolong the circulation time of the phages in the blood of rats.

[0064] In order to further confirm the time-delay effect of BCP1, we injected equal amounts of BCP1 and SC phages into rats respectively, and detected t...

Embodiment 3

[0068] Example 3: Verification that the time-delay function of phage has sequence specificity

[0069]During the screening process, we found that as the phages were enriched round after round, the frequency of phages containing RGD sequences increased continuously. Therefore, in order to determine the importance of RGD in the phage display peptide sequences obtained in Example 1, we constructed Three BCP1 phages were identified, as follows:

[0070] TB1: CRNHDMGAC (SEQ ID NO: 6, disrupted BCP1 sequence);

[0071] TB2: CNAAGAMHC (SEQ ID NO: 7, mutated RGD to AGA);

[0072] TB3: CAARGDAAC (SEQ ID NO: 8, RGD retained, remaining four amino acids mutated to A).

[0073] The 1×10 11 A TB1, TB2, TB3, BCP1, and SC phage were injected into different rats respectively, or mixed with REW 1:1 and injected into the same rat, and the number of phages in the blood at different time points was detected, and the results were as follows: Figure 5 shown.

[0074] Figure 5 The results sho...

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Abstract

The invention discloses a polypeptide for prolonging the blood circulation time of a bacteriophage, the bacteriophage, a method for screening the polypeptide and application of the polypeptide and thebacteriophage. The polypeptide is carried by the bacteriophage, and the bacteriophage has a function of prolonging the blood circulation time. The polypeptide, the bacteriophage, the method and the application have the advantage of important significance in bacteriophage therapy.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a polypeptide that prolongs the blood circulation time of bacteriophages. Background technique [0002] Phage therapy is a treatment method that uses phages to fight bacterial infections and has been used in clinical practice since the beginning of the 20th century. For example, in 1919, d'Herelle attempted to use phages to treat avian influenza (Salmonella), rabbit dysentery (Shigella) and bovine hemorrhagic sepsis (Pasteurella), the first attempts to use phage therapy. In 1921, Richard Bruynoghe and Joseph Maisin used phages to treat human skin diseases caused by Staphylococcus aureus. In 1932, at the request of the British government, d’Herelle went to the cholera endemic area in India to try to use bacteriophages for prevention and control, and quickly controlled the spread of the epidemic and effectively contained the second outbreak of the epidemic. In the same year...

Claims

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Application Information

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IPC IPC(8): C07K7/06C12N15/11C12N7/00C12Q1/70C12Q1/6869A61K35/76A61P31/04C12R1/92
CPCA61P31/04A61K35/76C12N7/00C12Q1/6869C07K7/06C12N2795/00021
Inventor 温龙平金佩佩沙瑞张云娇
Owner SOUTH CHINA UNIV OF TECH
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