Preparation method of antigen and adjuvant co-delivery nano-vaccine applied to liver cancer immunotherapy

An immunotherapy and nano-vaccine technology, applied in the field of biomedicine, can solve the problems of high cost of tumor vaccine raw materials, unfavorable large-scale processing and production, low immunotherapy effect, etc., and achieves easy repeatability of the preparation method, good application prospects, and enhanced immunotherapy. effect of effect

Active Publication Date: 2019-12-20
INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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  • Description
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Problems solved by technology

[0008] The present invention designs an antigen and adjuvant co-delivery nano-vaccine for immunotherapy of liver cancer and its preparation method. The technical problem it solves is that the existing tumor vaccine The immunotherapy effect on tumors is low, and the cost of raw materials for tumor vaccines is high, and the preparation method is complicated, which is not conducive to large-scale processing and production

Method used

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  • Preparation method of antigen and adjuvant co-delivery nano-vaccine applied to liver cancer immunotherapy
  • Preparation method of antigen and adjuvant co-delivery nano-vaccine applied to liver cancer immunotherapy
  • Preparation method of antigen and adjuvant co-delivery nano-vaccine applied to liver cancer immunotherapy

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preparation example Construction

[0034] The preparation method of antigen and adjuvant co-delivery nano vaccine applied to liver cancer immunotherapy comprises the following steps:

[0035] Step 1, the GPC3 127-136 Aqueous solution and polyethylenimine aqueous solution are mixed to obtain PC3 127-136 Mix solution with polyethyleneimine, then mix sodium alginate aqueous solution and CpG aqueous solution to obtain sodium alginate and CpG mixed solution, wherein GPC3 127-136 The concentration of aqueous solution is 0.5 mg / ml~4 mg / ml; the concentration of polyethyleneimine aqueous solution is 0.5 mg / ml~3 mg / ml; the concentration of CpG aqueous solution is 0.5 mg / ml~4 mg / ml; sodium alginate The concentration of the aqueous solution is 0.5 mg / ml~2 mg / ml; GPC3 127-136 The mass ratio between the aqueous solution and the polyethyleneimine aqueous solution is 1:0.05-0.5; the mass ratio between the sodium alginate aqueous solution and the CpG aqueous solution is 1:0.8-1.5.

[0036] Step 2: Add the mixed solution of s...

Embodiment 1

[0038] Preparation of Nanovaccine:

[0039] GPC3 127-136Aqueous solution (1mg / ml, 0.5ml) was mixed with PEI aqueous solution (0.5mg / ml, 0.2ml), then ALG aqueous solution (0.5mg / ml, 0.5ml) was mixed with CpG aqueous solution (0.5mg / ml, 0.5ml), Under the condition of stirring, add the mixed solution of ALG and CpG to GPC3 dropwise 127-136 and PEI mixed solution, stirred for 5 minutes to obtain antigen and adjuvant co-delivery nano-vaccine.

[0040] like figure 1 As shown, the particle size distribution of the nano-vaccine obtained in this example is relatively uniform, with an average particle size of 111.9 nm.

[0041] Evaluation of nano-vaccine promoting DC maturation:

[0042] Collect the DC of immature on the sixth day, press 10 6 cells per well, spread DC on 24-well plate, and add nano-vaccine, GPC3 after 2 hours of attachment 127-136 The concentration was 10 micrograms per milliliter, and the culture was continued for 48 hours. The cells were collected in a centrifu...

Embodiment 2

[0045] Preparation of Nanovaccine:

[0046] GPC3 127-136 Mix aqueous solution (2mg / ml, 1ml) and PEI aqueous solution (2mg / ml, 0.3ml), then mix ALG aqueous solution (2mg / ml, 0.1ml) and CpG aqueous solution (2mg / ml, 0.1ml), under stirring conditions, Add ALG and CpG mixed solution dropwise to GPC3 127-136 and PEI mixed solution, stirred for 2 minutes to obtain antigen and adjuvant co-delivery nano-vaccine.

[0047] like image 3 As shown, the particle size distribution of the nano-vaccine obtained in this example is relatively uniform, with an average particle size of 173.7nm.

[0048] Evaluation of nano-vaccine promoting DC cytokine secretion:

[0049] Collect the DC of immature on the sixth day, press 10 6 cells per well, spread DC on 24-well plate, and add nano-vaccine, GPC3 after 2 hours of attachment 127-136 The concentration was 10 micrograms per milliliter, and the culture was continued for 48 hours. The cells were collected in a centrifuge tube by gentle blowing, ...

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Abstract

The invention relates to a preparation method of an antigen and adjuvant co-delivery nano-vaccine applied to liver cancer immunotherapy. The preparation method comprises the steps as follows: step 1,mixing a GPC3127-136 aqueous solution and a polyethylenimine (PEI) aqueous solution to obtain GPC3127-136 and PEI mixed solution, and mixing a sodium alginate aqueous solution and a CpG aqueous solution to obtain a sodium alginate and CpG mixed solution; and step 2, dropwise adding the sodium alginate and CpG mixed solution to the GPC3127-136 and PEI mixed solution under the stirring condition, and performing stirring continuously to obtain the antigen and adjuvant co-delivery nano-vaccine applied to liver cancer immunotherapy.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a preparation method of a nano-vaccine co-delivered with an antigen and an adjuvant for liver cancer immunotherapy. Background technique [0002] The incidence and mortality of liver cancer are increasing year by year, seriously endangering human health and safety. Immunotherapy for liver cancer is a treatment method to control and eliminate tumors by reactivating the body's normal anti-tumor immune response. [0003] Tumor vaccines play an important role in tumor immunotherapy. Tumor vaccines are processed and processed by antigen-presenting cells in vivo, and present antigen information to T cells, thereby stimulating the body's immune response. Dendritic cells (DC) are the most powerful professional antigen-presenting cells known in vivo, and are the central link in the initiation, regulation and maintenance of immune responses. [0004] Finding effective antigens is crucial for t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/39A61K47/69A61K47/32A61K47/34A61P35/00
CPCA61K39/001174A61K39/39A61K47/6933A61K47/6939A61P35/00A61K2039/55561A61K2039/622A61K2039/844Y02A50/30
Inventor 张闯年王晓莉孔德领孙洪范裴萌月徐蓉
Owner INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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