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Immunogenic molecules

An immunogenic and molecular technology, applied in the field of immunology, can solve problems such as ineffectiveness and ineffectiveness of full-length proteins

Inactive Publication Date: 2008-03-26
COUNCIL OF THE QUEENSLAND INST OF MEDICAL RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, use of these strategies in viral vaccination or therapy generally results in an evolutionary switch in the viral genome where this response is nullified
Delivery strategies are also limited because full-length proteins containing CTL epitopes do not efficiently enter the MHC class I processing pathway

Method used

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  • Immunogenic molecules
  • Immunogenic molecules
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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0182] Materials and Methods

[0183] Reagent

[0184] Unless otherwise specified, chemical reagents are of analytical grade or equivalent. N,N'-dimethylamide (DMF), piperidine, trifluoroacetic acid (TFA), O'-benzotriazole-N,N,N',N'-tetramethylurea cationic hexafluorophosphate (HBTU), 1-Hydroxybenzotriazole (HOBt), Diisopropylethylamine (DIPEA) and Diisopropylcarbodiimide (DIPCDI) were from Auspep Pty in Melbourne, Australia and Sigma- Aldrich Pty Company. Dichloromethane (DCM) and diethyl ether were purchased from MerckPty (Kilsyth, Australia). Phenol and triisopropylsilane (TIPS) are from Aldrich (Milwaulke, WI), trinitrobenzene methanesulfonic acid (TNBSA) and diaminopyrimidine (DMAP) are from Fluka; 1,8-diazabicyclo[ 5.4.0] Undec-7-ene (DBU) is from Sigma, palmitic acid is from Fluka. The solid supports TentaGel S RAM and TentaGel S Am were from Rapp Polymere GmbH, Tubingen, Germany. O-(N-Fmoc-2-aminoethyl)-O'-(2-carboxyethyl)-undecaethylene glycol (Fmoc-PEG) was obtained from...

Embodiment 2

[0210] Synthesis of four different types of lipidated HEL (lysozyme) proteins

[0211] Four different lipidated HELs were prepared by coupling the four lipid moieties listed in Figure 1 to the egg lysozyme (HEL) protein. Figure 3 shows a schematic diagram of these four lipidated proteins.

[0212]Lipidation 1HEL (thioether) and lipidation in lipidated HEL protein 2 HEL (thioether) is prepared by derivatizing HEL with MCS and then chemically selectively connecting the sulfhydryl group of structure A to form a thioether bond between the protein and the lipid assembly. Their difference lies in lipidation 2 HEL (thioether) has two copies of structure A. Lipidation of branches 2 HEL has only one copy of lipid component per protein molecule, but because of the bivalent nature of structure B, there are two copies of pam2cys per protein. This makes it more hydrophobic and elution by HPLC is much delayed.

[0213] To prepare lipidation 1 HEL (disulfide), HEL is modified with iso(yl) bifunc...

Embodiment 3

[0215] Evaluation of the immunogenicity of lipidated insulin

[0216] The insulin is lipidated by the following procedure. 10mg of bovine insulin was dissolved in 400μl of 0.5M phosphate buffer (pH 7.9) containing 6M guanidine hydrochloride and 400μl of 0.02M phosphate buffer (pH 7). 200μl of acetonitrile solution containing 3.25mg of N-succinimidyl 6-maleimidohexanoate (MCS) was added to this solution. After 3 hours of action, the MCS-modified insulin was treated by semi-preparative HPLC purification. 3.3 mg MCS modified insulin and 5 mg Pam2CysSer(Lys)8Cys were dissolved in 500 μl acetonitrile and 500 μl water. The reaction mixture was left at room temperature for 48 hours. Two lipidated insulin compounds were separated by semi-preparative HPLC. Using mass spectrometry to analyze these components showed that two different lipidated insulins were obtained, and the number of lipid moieties attached to each protein molecule was different. Pam2Cys 2 -Insulin has two copies of Pam pe...

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Abstract

The present invention relates generally to the field of immunology and more particularly to molecules capable of stimulating a cellular immune response. More particularly, the present invention provides self-adjuvanting immunogenic molecules capable of stimulating an immune response to epitopes of a polypeptide irrespective of a subjects HLA type. The present invention further contemplates methods for the production and use of the self- adjuvanting immunogenic molecules and compositions comprising same useful in the vaccination of subjects against specific polypeptides.

Description

Technical field [0001] The present invention relates generally to the field of immunology, and more specifically to molecules capable of stimulating cellular immune responses. More specifically, the present invention provides a self-adjuvant (self-adjuvanting) immunogenic molecule that can stimulate an immune response against a polypeptide epitope regardless of the HLA type of the subject. The present invention also relates to methods for producing and using self-adjuvant immunogenic molecules and compositions comprising the self-adjuvant immunogenic molecules, which can be used for vaccination of subjects against specific polypeptides. Background technique [0002] Any prior art mentioned in this specification is not, and should not be regarded as an endorsement or suggestion in any form that the prior art forms part of Australia’s common knowledge. [0003] Immunotherapy and vaccination are used to prevent or treat a wide range of diseases, such as infectious diseases and certa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K38/04A61K38/17
CPCA61K39/292A61K39/00C07K2319/00A61K39/245A61K2039/6018A61K39/0008C12N2730/10134A61K2039/543C12N2710/16634A61K39/12A61P35/00A61P35/02C07K19/00
Inventor D·C·杰克森曾伟光
Owner COUNCIL OF THE QUEENSLAND INST OF MEDICAL RES