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Recombinant Inactivated Virus Vector Vaccine

一种重组疫苗、病毒载体的技术，应用在病毒/噬菌体、疫苗、病毒等方向，能够解决稳定性有限、产业方面复杂、乳液没有优势等问题，达到减少排毒、减少病毒的传播、减少爆发流行的风险的效果

Inactive Publication Date: 2011-12-14
LAB AVI MEX S A DE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0028] Likewise, recombinant vaccines have not yet been used in an inactivated form, as this would mean that the concentration of the viral vector would be 100 times higher than that required for normal virus (10 times higher than recombinant live virus), which would be very industrially critical. complex
As a result, these recombinant live virus vaccines have never been used as emulsions in general because of their limited stability and because emulsions have no advantage in this regard due to the active characteristics of live viral vectors

Method used

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  • Recombinant Inactivated Virus Vector Vaccine
  • Recombinant Inactivated Virus Vector Vaccine
  • Recombinant Inactivated Virus Vector Vaccine

Examples

Experimental program
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Effect test

Embodiment 1

[0076] Generation of Newcastle disease-LaSota vector

[0077] In order to clone the genome of Newcastle disease virus LaSota strain and thus prepare the virus vector, the intermediate vector "pNDV / LS" was first produced. Total viral RNA of Newcastle disease LaSota strain was extracted by triazole method. cDNA (complementary DNA) was synthesized from the purified RNA of the viral genome using the previously purified total RNA as a template. To clone all genes of the Newcastle disease genome (15,183 base pairs (bp)), seven fragments with "overlapping" ends and cohesive restriction sites were amplified by PCR. Fragment 1 (F1) comprises nucleotides (nt) 1-1755, F2 comprises nt 1-3321, F3 comprises nt 1755-6580, F4 comprises nt 6,151-10,210, F5 comprises nt 7,381-11,351 , F6 contains nt 11,351-14,995 and F7 contains nt 14,701-15,186. In the cloning vector pGEM-T, standard ligation techniques were used to assemble the seven fragments to reconstruct the Newcastle disease LaSota ge...

Embodiment 2

[0079] Cloning of HA Gene from VIA Subtype H5N2435 Strain 435

[0080] The total virus RNA was extracted by triazole method in order to clone the HA gene of VIA 435 strain. The purified total RNA was then used to synthesize cDNA (complementary DNA), and the HA gene of the IA virus was amplified with specific oligonucleotides by PCR technique. Then, the HA gene of 435 was inserted into the pGEM-T vector using standard cloning techniques to obtain a plasmid: p-GEMT-435.

Embodiment 3

[0082]Cloning the IA 435HA gene with the SacII site of the pNDV / LS vector to make a plasmid: pNDV / LS-435

[0083] A. Prepare the intermediate vector pSacIIGE / GS:

[0084] The intermediate vector pSacIIGE / GS was constructed to introduce the transcribed sequence GE / GS of Newcastle disease at the 5' end of the HA 435 gene. The method was to perform PCR initial amplification of the GE / GS sequence using the Newcastle disease genome as a template, and then these sequences were Insert into pGEM-T.

[0085] B. Subcloning the HA gene into the pSacIIGE / GS vector:

[0086] Plasmid pGEMT-435 was digested with Hpal-Ndel and cloned into pSacIIGE / GS to obtain plasmid pSacIIGE / GS-HA435.

[0087] C. Subcloning GE / GS-HA435 into pNDV-LS vector

[0088] Two plasmids: pSacIIGE / GS-HA435 and pNDV / LS were digested with SacII, the digested products were purified, the GE / GS-HA435 region was purified and inserted into the SacII site of pNDV / LS, and an infectious clone was obtained: pNDV / LS- 43...

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Abstract

The present invention describes a vaccine, which comprises an inactivated viral vector and a pharmaceutically acceptable vehicle, adjuvant or excipient, and an exogenous nucleotide sequence encoding a target disease is inserted into the vector, and the vaccine is obtained by using A viral vector titer similar to that required for a live virus vaccine based on the same viral vector provides the desired protection against the target disease. Primarily, the present invention describes viral vectors of paramyxoviruses or adenoviruses.

Description

technical field [0001] The present invention relates to techniques for the prevention and treatment of diseases, especially poultry diseases, and more particularly, to recombinant vaccines containing inactivated viral vectors and pharmaceutically acceptable vehicles, adjuvants or excipients, said vectors having an inserted coding An exogenous nucleotide sequence of a protein that has antigenic activity for a disease. Background of the invention [0002] It is well known that vaccines against viral pathogens are formulated from the corresponding viruses, which are isolated for the production of vaccines to be administered to animals or humans through diverse formulations. [0003] On the one hand, some vaccine products are made from intact live viruses that show low pathogenicity in the wild, or from viruses that have been attenuated in the laboratory and have reduced pathogenicity. Provides protection against highly pathogenic strains of the same species. [0004] For exam...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/145A61K39/00
CPCA61K2039/5256A61K2039/545C12N2710/10343C12N2760/16134A61K39/145A61K2039/5252A61K2039/552A61K2039/55566A61K2039/70C12N2760/18134C12N2760/18143C12N2770/20034A61K39/12A61P31/04A61P31/12A61P31/14A61P31/16A61P31/20A61P37/04A61K2039/54A61K2039/55C12N7/00C12N2710/10043C12N2760/18163C12N2790/00034
Inventor B.洛扎诺-杜伯纳德D.萨法蒂-米兹拉西J.A.苏亚雷斯-马丁内兹M.J.盖伊-古蒂雷兹E.索托-普里安特
Owner LAB AVI MEX S A DE
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