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Chicken marrow-derived dendritic cell (DC) targeting peptide AH and application thereof

A technology of dendritic cells and targeting peptides, applied in the field of biomedicine, can solve the problems of poor binding effect of poultry-derived DCs, achieve the effects of reducing degradation rate and side effects, improving immune efficiency, and being easy to synthesize

Active Publication Date: 2018-07-13
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although existing mammalian DC targeting peptides can bind to DCs from multiple species to varying degrees, they are less effective on avian DCs
Poultry vaccines have not yet involved targeted drug delivery, and there have been no reports on poultry-derived DC targeting peptides

Method used

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  • Chicken marrow-derived dendritic cell (DC) targeting peptide AH and application thereof
  • Chicken marrow-derived dendritic cell (DC) targeting peptide AH and application thereof
  • Chicken marrow-derived dendritic cell (DC) targeting peptide AH and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation, cultivation and identification of chicken bone marrow-derived dendritic cells

[0027] 1. Isolation and culture of chicken bone marrow-derived dendritic cells

[0028] Chickens about 30 days old were sacrificed and soaked in 75% ethanol for 10 minutes. The femur and tibia were taken out under aseptic conditions, and PBS containing 2% double antibody (pH 7.2) was sucked with a syringe, and the bone marrow was washed until the marrow cavity became white; For the washing solution, wash twice with PBS containing 2% double antibody, centrifuge at 1000r / min for 5-10min each time, and discard the supernatant; collect the precipitate and resuspend the precipitated cells with PBS, and then dilute the bone marrow cell suspension at a ratio of 1:1. Slowly add to the centrifuge tube containing Histopaque-1119, centrifuge at 2500r / min for 30min; collect the white misty cells at the interface in the middle, resuspend in PBS, centrifuge at 1500r / min for 10min, dis...

Embodiment 2

[0036] Example 2. Preliminary detection of phage display 12-peptide library subtractive screening for targeting peptides and their affinity for DC

[0037] 1. Phage display 12-peptide library subtractive screening targeting peptides

[0038] The screening process is divided into four rounds, as follows:

[0039] First round: place 2 x 10 11 The phage display 12 peptide library (PhD-12) library of pfu was added to the chicken bone marrow derived dendritic cells (5 × 10 5 / mL), and incubated at 4°C for 30min, centrifuged at 2000rpm for 3min, resuspended the cell pellet in PBS containing 1% bovine serum albumin (BSA) and 0.05% Tween 20, and repeated washing 3 times; adding 1mL Buffer solution (0.2mol / L glycine-hydrochloric acid, pH=2.2, 1mg / mL bovine serum albumin), mix lightly at room temperature for 10 minutes, then add 50μl Tris-HCl (pH value 9.1) for neutralization; centrifuge, and the supernatant is The phage eluate was bound to the surface of the membrane, and the eluate...

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Abstract

The invention discloses a chicken marrow-derived dendritic cell (DC) targeting peptide AH and an application thereof. The amino acid sequence of the chicken marrow-derived DC targeting peptide AH is shown as SEQ ID NO.1. A chicken marrow-derived DC dominant binding peptide AH is sieved by using a phage random peptide library and a whole cell differential screening technology, the peptide can be relatively strongly bound to a chicken marrow-derived DC to show good targeting properties and specificity, and therefore, the targeting peptide can be used as a lead molecule of a drug such as a gene leading carrier to lay the foundation for specific small molecular therapy of a cell or can be combined with a vaccine to increase the immunization efficiency of an avian vaccine and promote organismsto generate immunities in advance.

Description

technical field [0001] The invention relates to a chicken bone marrow-derived dendritic cell targeting peptide and its application, belonging to the technical field of biomedicine. Background technique [0002] The rapid development of immunology and molecular biology has promoted the development of new vaccines, which use the interaction between immune cells and vaccine antigens to optimize the body's response to speed up the immune response and improve the specificity of acquired immunity. In recent years, targeted therapy and prevention have become research hotspots in the field of medical biotechnology, and have been applied to the treatment of human tumors. Among them, biological immunotherapy targeting dendritic cells can effectively deliver drugs to the target site, Reduce the dose and frequency of administration, improve the therapeutic effect of the drug, and reduce adverse reactions. Dendritic cells (DCs) are the most powerful professional antigen-presenting cells...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12N15/12A61K47/42
CPCA61K47/42C07K7/08
Inventor 李一经徐义刚王丽唐丽杰马孙婷
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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