Recombinant beta-lactamase, usable as carrier molecule in immuno

Drug – bio-affecting and body treating compositions – Whole live micro-organism – cell – or virus containing – Genetically modified micro-organism – cell – or virus

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Details

536 232, 536 234, 4352523, 43525411, 4352543, 4353201, 530350, C07H 2104, C12N 121, C12N 119, C12N 1563

Patent

active

058304576

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BRIEF SUMMARY
This application is a 371 of PCT/FR93/00151 Feb. 12, 1993.
The object of the present application is novel agents for the preparation of immunogenic compositions and preferably protective vaccinating compositions available in the form of "live vaccines", particles or molecules which can be administered to man or animals.
One of the objectives of the present invention is to suggest agents for the development of immunogenic compositions capable of triggering in man or animals a cellular and/or humoral immune response (through the intermediary of antibodies) against antigenic determinants and in particular against epitopes characteristic of different pathogenic agents. More generally, the invention suggests novel agents to trigger or promote an immune response against any specific epitope to produce antibodies or a cellular immune response, and in particular when the antigenic determinants are of the hapten type and consequently incapable of triggering this immune response by themselves. The invention is also of interest for the purpose of promoting an immune response capable of being conferred by an antigen, and in particular of enhancing the level or nature of the protection.
In this connection the invention describes novel molecules capable of being used as vectors of the antigenicity of different epitopes both in immunogenic compositions containing the hybrid (recombinant) molecules thus formed or in live vaccines.
Different factors can determine the selection of a molecule, in particular of a protein capable of behaving as carrier protein (also called vector) for the purpose of conferring or improving the antigenicity of an epitope heterologous with respect to this molecule. For example, the size parameters of the vector as such and its size with respect to that of the epitope which it will contain as well as the size of the epitope as such must be taken into account. Other constraints for the development of vaccines, live or not, implicating carrier molecules are, for example, the antigenicity of the carrier molecule, its toxicity for a cell host in which it would be produced or for the subject to whom it would be administered. It is also advisable to determine the potential sites of insertion for the epitope and--a matter of some importance when this modified protein is used in the context of the preparation of a live vaccine--its capacity to be exported, even secreted by the host which produces it in order to be accessible to the immune system of the subject to whom the vaccine is administered.
In the context of the present invention the inventors were interested in proteins of the beta-lactamase family, certain properties of which have been studied up to now in a context different from that of the present invention.
The beta-lactamases are enzymes arranged in different classes as a function of their enzymatic activity on different substrates. They are produced by different organisms and in particular by bacteria for example bacteria of the E. coli, Staphylococcus aureus type, or also by mycobacteria. These enzymes have been studied for their capacity to protect bacteria against the lethal effects of antibiotics of the beta-lactam type, and hence their capacity to confer on certain bacteria resistance to different antibiotics. It is in this context of the study of the resistance of bacteria to antibiotics that several authors have published articles describing the purification of certain beta-lactamases.
For example, Choubey et al. have published results describing the purification to homogeneity of a beta-lactamase from Mycobacterium smegmatis (Microbiology, 1986, vol. 13: 171-175). Other authors have published data relating to the characterization of the beta-lactamase produced in a strain of Mycobacterium fortuitum(J. Amicosante et al., Biochem. J., 1990, 271: 729-734; L. Fattorili et al., Antimicrobial Agents and Chemotherapy, September 1991, p. 1760-1764). In this second publication, the authors point out that the beta-lactamases have been described as being periplasmic enzymes in Gram-negative ba

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