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Nano vaccine of LAMP-2 epitope peptide and preparation method of nano vaccine

A technology of LAMP-2, 1.LAMP-2, applied in the field of biomedicine, can solve problems such as failure to find, achieve the effect of enhancing drug efficacy and eliminating spontaneous immune response

Inactive Publication Date: 2014-02-05
THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In ANCA-associated vasculitis, the confirmed ANCA-associated target antigens include neutrophil myeloperoxidase (MPO) and protease 3 (protease3, PR3), but it has not been found that the infection is related to MPO or PR3 direct evidence of

Method used

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  • Nano vaccine of LAMP-2 epitope peptide and preparation method of nano vaccine
  • Nano vaccine of LAMP-2 epitope peptide and preparation method of nano vaccine
  • Nano vaccine of LAMP-2 epitope peptide and preparation method of nano vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 loaded with LAMP-2 epitope peptide P 331-341 Preparation of Nanovaccine

[0046] Weigh 5mg of LAMP-2 epitope peptide P with electronic balance 331-341 (purity>95%, Hangzhou Zhongpei Biochemical Co., Ltd.) and 15mg poly(lactic-co-glycolic acid) (LA:GA=50 / 50, sigma), put in a 10ml centrifuge tube, add 0.6ml dichloromethane, and vortex to mix Stir it into a homogeneous mixture, add 6ml of phosphate buffer (pH 6.5) containing 1% polyvinyl alcohol, and place it under the ultrasonic probe for 20s (power 90W, time 20s, pause interval 1s every 3s). Pour the emulsion into 20ml of phosphate buffer solution (pH 6.5) containing 0.3% polyvinyl alcohol, put it into a stirring bar, and stir with a magnetic stirrer for 3 hours at room temperature to evaporate the organic solvent. Pour the emulsion after the organic solvent has evaporated into an EP tube, put it into a centrifuge tube and centrifuge (15000rpm, 10min), pour off the supernatant, add double-distilled water, and...

Embodiment 2

[0047] Example 2 is loaded with LAMP-2 epitope peptide P 41-49 Preparation of Nanovaccine

[0048] Weigh 5mg of LAMP-2 epitope peptide P with electronic balance 41-49 (purity>95%, Hangzhou Zhongpei Biochemical Co., Ltd.) and 15mg poly(lactic-co-glycolic acid) (LA:GA=50 / 50, sigma), put in a 10ml centrifuge tube, add 0.6ml dichloromethane, and vortex to mix Stir it into a homogeneous mixture, add 6ml of phosphate buffer (pH 6.5) containing 1% polyvinyl alcohol, and place it under the ultrasonic probe for 20s (power 90W, time 20s, pause interval 1s every 3s). Pour the emulsion into 20ml of phosphate buffer solution (pH 6.5) containing 0.3% polyvinyl alcohol, put it into a stirring bar, and stir with a magnetic stirrer for 3 hours at room temperature to evaporate the organic solvent. Pour the emulsion after the organic solvent has evaporated into an EP tube, put it into a centrifuge tube and centrifuge (15000rpm, 10min), pour off the supernatant, add double-distilled water, and ...

experiment example 3

[0049] Evaluation of Experimental Example 3 Nano Vaccine

[0050] (1) Morphological observation of the prepared nano-vaccine

[0051] Dilute the nanoparticle suspension washed in Examples 1 and 2 and evenly spread it on the mica sheet. After natural drying, observe the morphology with a scanning electron microscope (S-3400N electron microscope). The test results show that the nanoparticles prepared by the emulsified solvent volatilization method have good spheroidity, good shape and uniform particle size. like figure 1 Shown is the scanning electron micrograph of the nano-vaccine prepared in Example 2.

[0052] (2) Particle size test of the prepared nano-vaccine

[0053] The nanoparticles washed in Examples 1 and 2 were diluted with double distilled water and loaded into the sample cell of a particle size analyzer (Malvern Company) to measure the particle size distribution. The test results show that the particle size of the nanoparticles loaded with epitope peptide is bet...

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Abstract

The invention belongs to the field of biological medicines, and relates to a nano vaccine of LAMP-2 epitope peptide and a preparation method of the nano vaccine. The nano vaccine comprises an effective amount of polypeptide with the amino acid sequence as shown in SEQ IN DO:1, or a polypeptide mixture with the amino acid sequence as shown in SEQ IN DO:1 and SEQ IN DO:2, the nano vaccine can be prepared through the synergistic effect of a polylactic acid-glycolic acid copolymer with the polypeptide, and the nano vaccine can be applied to preparation of a nano vaccine for anti-neutrophile granulocyte cytoplasm antibody relevancy vasculitis and anti-LAMP-2 antibody relevancy pauci-immune complex crescent glomerulonephritis. Due to adoption of the nano vaccine, spontaneous immune response of autoantigen characteristics is eliminated, and immune response to other antigens is not interfere, and a novel method is provided for treatment on anti-neutrophile granulocyte cytoplasm antibody relevancy vasculitis and anti-LAMP-2 antibody relevancy pauci-immune complex crescent glomerulonephritis.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a nano-vaccine of LAMP-2 antigen epitope peptide, as well as its specific application form and preparation method. Background technique [0002] Pauci-immune crescentic glomerulonephritis (PICGN) is a local manifestation of antineutrophil cytoplasmic autoantibodies (ANCA)-associated vasculitis, characterized by hematuria, proteinuria, edema, high Blood pressure is the predominant clinical manifestation and can lead to rapidly progressive, irreversible renal failure. It is characterized by little or no immune complex deposition on the glomerular capillary wall, glomerular capillary loop necrosis and crescent formation, and neutrophils directly cause damage to the microvascular endothelium and glomerulus. The progression of the disease is dangerous, with a mortality rate as high as 15% one year after diagnosis. Even with timely and powerful immunosuppressive treatment, the mortality rate w...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K9/14A61K47/34A61P9/14A61P13/12A61K39/39
Inventor 张静波尹世伟唐莎张莹
Owner THE SECOND AFFILIATED HOSPITAL ARMY MEDICAL UNIV
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