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Liposomal formulations

A technology of liposomes and compositions, which is applied in liposome delivery, antibody medical components, viruses/phages, etc., and can solve problems such as reducing liposome efficiency

Inactive Publication Date: 2014-04-23
MOLECULAR EXPRESS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Liposomes are rapidly cleared by the reticuloendothelial system and macrophages, reducing the efficacy of liposomes as vaccines

Method used

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  • Liposomal formulations
  • Liposomal formulations
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Examples

Experimental program
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Effect test

preparation example Construction

[0218] The preparation of liposomes is well known in the art. In general, liposomes have been prepared by a number of different techniques, including ethanol infusion (Batzri et al., Biochem. Biophys. Acta. 298:1015, 1973); ether infusion (Deamer et al., Biochem. Biophys. Acta. 443 Schieren et al., Biochem.Biophys.Acta.542:137,1978); Detergent removal (Razin, Biochem.Biophys.Acta.265:241972); Solvent evaporation (Matsumato et al., J.Colloid Interface Sci .62:149,1977); evaporation of organic solvents (REV) from aqueous emulsions of chloroform (Szoka Jr. et al., Proc.Natl.Acad.Sci.USA,75:4194,1978); Extrusion of MLV or EUV (Olson et al., Biochem. Biophys. Acta. 557:9, 1979); freeze-thawing of phospholipid mixtures (Pick, Arch. Biochem. Biophys., 212:186, 1981) and sonication and homogenization. By convention, liposomes are classified by size and a 3-letter acronym is used to designate the type of liposome in question. Multilamellar vesicles are often designated "MLV". Small ...

Embodiment 1

[0238] Embodiment 1. Influenza A virus materials and methods

[0239] A. IAVM2e1-HDpET28a vector

[0240] The IAVM2e1-HD pET28a plasmid design has been described previously [Ernst et al., 2006].

[0241] B. IAVM2e1-HD pEV vector

[0242] Table 1. Peptide and oligonucleotide sequences of the different IAVM2e1 sequences evaluated

[0243]

[0244] To generate the IAVM2e1-HD protein, a 5' oligonucleotide primer (Integrated DNA Technologies) encoding the IAVM2el gene containing a Bgl II restriction enzyme site and an enterokinase (EK) site was synthesized. To generate the LDR-IAVM2e1-HD protein, a 5' oligonucleotide primer (Integrated DNA Technologies) encoding the IAVM2e1 gene containing the Bgl II restriction enzyme site, the EK site and the LDR sequence of the pET28a vector was synthesized. Similarly, a 3' oligonucleotide primer encoding a hydrophobic domain gene containing an Xho I restriction enzyme site was synthesized. To generate the IAVM2e1-HD gene, primers were us...

Embodiment 2

[0291] Example 2. Results

[0292] In this study, in addition to pET28a-IAVM2e1-HD, we constructed four new IAVM2e1-HD constructs: (i) pEVlIAVM2e1-HD with the above LDR sequence, (ii) pEVl-IAVM2e1(C16S,C18S) -HD, (iii) pEV1-IAVM2e1(C16S)-HD, (iv) pEV1-IAVM2e1(C18S)-HD and (v) pEV1-IAVM2e1(1-15)-HD. All new plasmids were sequenced and confirmed to be in frame and contain the correct sequence. PCR products for each IAVM2e1-HD construct indicated that each PCR product corresponded to the expected size of the corresponding IAVM2e1-HD insert. The plasmid was transformed into E. coli strain C-43 and expressed. Small scale expression studies were performed to confirm that the construct expressed the protein. Unlike the original pET28a-IAVM2e1-HD construct, the pEV-containing IAVM2e-HD construct showed high levels of protein expression compared to the corresponding uninduced controls. To confirm expression of the target M2e protein, Western blot analysis was performed. The IAVM2e...

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Abstract

The present invention relates to liposomal vaccine compositions, methods for the manufacture thereof, and methods for the use thereof to stimulate an immune response in an animal. These compositions comprise liposomes formed from dimyristoylphosphatidylcholine ("DMPC"); either dimyristoylphosphatidylglycerol ("DMPG") or dimyristoyltrimethylammonium propane ("DMTAP") or both DMPC and DMTAP; and at least one sterol; and an antigenic polypeptide comprising a first polypeptide sequence, and a second polypeptide sequence heterologous the first polypeptide sequence which comprises a transmembrane sequence from a membrane protein, said transmembrane sequence having a number of residues sufficient to cross a lipid bilayer, at least nine contiguous residues of which are predicted to form an alpha helix having a hydrophobicity score of about 0.7 or greater.

Description

[0001] Related applications [0002] This application claims priority to US Provisional Patent Application No. 61 / 479,302, filed April 26, 2011, which is hereby incorporated in its entirety, including all tables, drawings, and claims. Background of the invention [0003] Statement Regarding Federally Funded Research [0004] Under the terms of National Institute of Allergy and Infectious Diseases (NIH) grant numbers U01AI074508 and 1R43AI078654, the U.S. Government has a paid-up license to this invention and the right, under limited circumstances, to require the patent owner to license others on reasonable terms. [0005] The following discussion of the background of the invention is provided solely to assist the reader in understanding the invention and is not an admission that it describes or constitutes prior art to the invention. [0006] Liposomes are vesicles formed by one ("unilamellar") or multiple ("multilamellar") layers of phospholipids. Due to the amphiphilic nat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K9/127
CPCA61K9/127A61K9/1277A61K39/145A61K2039/55555A61K2039/6031C07K2319/03C07K2319/35A61K45/06A61K39/12A61K2039/543A61K2039/55572C12N2760/16134A61P37/04A61K38/16A61K47/42
Inventor G·藤井W·A·恩斯特J·阿德勒-摩尔L·维茨安
Owner MOLECULAR EXPRESS