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Nano-platform for overcoming vector epitope inhibition effect of vaccine

A carrier and carrier protein technology, applied in the direction of medical preparations containing active ingredients, antibody medical ingredients, non-active ingredients of polymer compounds, etc., can solve the problems of non-green synthesis methods, complicated production operations, and high costs, and achieve the synthesis process Environmentally friendly, non-toxic and harmless raw materials, small batch differences

Active Publication Date: 2018-12-18
XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] In order to solve the problem of "carrier inhibition effect", there are literatures using polylactic acid-glycolic acid (PLGA) and liposomes as materials for coating carrier proteins. The existence of this material does not completely inhibit the production of carrier protein antibodies, and the synthesis method is not green ( The synthesis process involves toxic organic solvents, etc.), and the synthesis process is complex and cumbersome, and the synthesis materials are complex (including a variety of lipid components and polymer materials)
[0003] In addition, there are literatures that use polyethylene glycol (PEG) to modify the carrier protein to reduce the "carrier inhibition effect", but this solution does not completely inhibit the production of carrier protein antibodies, and long-term use of PEG will also produce antibodies against PEG, resulting in the body's resistance to PEG Accelerated Clearance
[0004] In addition, there are also literatures that use plasmid transfection and prokaryotic expression to produce recombinant proteins fused with self-antigens and non-self-antigens to reduce the "carrier inhibition effect". However, this solution does not completely inhibit the production of carrier protein antibodies, and the production operation is complicated high cost

Method used

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  • Nano-platform for overcoming vector epitope inhibition effect of vaccine
  • Nano-platform for overcoming vector epitope inhibition effect of vaccine
  • Nano-platform for overcoming vector epitope inhibition effect of vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] 1) Reduction of MSA (in order to reduce albumin disulfide bonds to form sulfhydryl groups to facilitate reassembly of cross-links)

[0091] Reduction of MSA / KLH: Weigh 1.5mg of KLH, 10.5mg of MSA, dissolve in 2.4ml of carbonic acid buffer solution with pH=9.0, the final concentration of total protein is 5mg / ml, and call it solution A; weigh several mg of DTT, dissolve in In ionized water, the final concentration is 10 mg / ml, which is solution B; mix 62ul of solution B and solution A in a 2.5ml reaction bottle, place it on a flat magnetic stirrer, and react at 300r for 1h at room temperature, and it is solution C.

[0092] 2) Thermal polymerization to form MAS-KLH cross-linked nanocarriers (MKN) (using SDS to regulate particle size, thermal polymerization and stirring, etc. Make the protein form nanoparticles with good uniformity first, and in the process, some of the sulfhydryl groups have re-formed new disulfide bonds)

[0093] Thermal polymerization to form MKN:...

Embodiment 2

[0102] (1) Reduction of MSA / HSA (in order to reduce albumin disulfide bonds to form sulfhydryl groups to facilitate reassembly of cross-links)

[0103] Reduction of HAS / KLH: Weigh several mg of DTT, dissolve in deionized water, the final concentration is 10mg / ml, which is solution B; weigh 1mg of KLH, 9mg of HSA, dissolve in 2ml of phosphate buffer with pH=8.0, total The final protein concentration is 5mg / ml, which is solution D; mix 62ul of B solution and D solution in a 2.5ml reaction bottle, place it on a flat magnetic stirrer, and react at 300r for 1h at room temperature, and this is solution E.

[0104] (2) Thermal polymerization to form HSA-KLH cross-linked nanocarriers (HKN) (using SDS to control the particle size, thermal polymerization and stirring, etc. Use the protein to form nanoparticles with good uniformity, and in the process, some of the sulfhydryl groups have re-formed new disulfide bonds)

[0105] Thermal polymerization to form HKN: add water to the react...

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Abstract

The invention provides a nano-platform for overcoming the vector epitope inhibition effect of a vaccine. Albumins used as a raw material are reduced, thermopolymerizied, oxidized and concentrated, andthen are externally linked with an antigen epitope to obtain a target vaccine vector, wherein the reduction is carried out to reduce disulfide bonds in the albumins into sulfhydryl groups; the thermopolymerization is carried out to obtain nano-particles formed in an albumin self-assembling manner; and the oxidation is carried out to oxidize the sulfhydryl groups into the disulfide bonds. The nano-platform is a nano-vaccine vector having the advantages of completeness in inhibiting the production of vector protein antibodies, green and simple synthesis process, no involving of toxic chemicalsor complicated processes, low cost, small batch difference and great clinical transformation prospect.

Description

technical field [0001] The invention relates to the field of nano vaccines, in particular to a nano platform for overcoming the inhibitory effect of vaccine carrier epitopes. Background technique [0002] In order to solve the problem of "carrier inhibition effect", there are literatures using polylactic acid-glycolic acid (PLGA) and liposomes as materials for coating carrier proteins. The existence of this material does not completely inhibit the production of carrier protein antibodies, and the synthesis method is not green ( The synthesis process involves toxic organic solvents, etc.), and the synthesis process is complex and cumbersome, and the synthesis materials are complex (comprising a variety of lipid components and polymer materials). [0003] In addition, there are literatures that use polyethylene glycol (PEG) to modify the carrier protein to reduce the "carrier inhibition effect", but this solution does not completely inhibit the production of carrier protein an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/42A61K39/00
CPCA61K39/00A61K47/42
Inventor 何斌石学银季海英薛晓梅洪婷杜健儿
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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