Hybrid cells producing an antigen characteristic of the hepatiti

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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435 693, 435 702, C12N 1506

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active

051127482

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BRIEF SUMMARY
The invention relates to hybrid cells which are effective in the production of a polypeptide having the immunologic and, preferably also, immunogenic, properties of an antigen of the hepatitis B virus, preferably of an antigen of the envelope of this virus, more particularly of the antigen HBsAg or of a fragment of it, this polypeptide being coded by a cloned or clonable sequence of DNA in this cellular hybrid. The invention equally relates to a process for the production of these cellular hybrids and to their application to the production of the said polypeptide.
The cellular hybrids of the present invention enter into the context of those which have already been described in the parent application, Ser. No. 06/800,080. It will be recalled that the cellular hybrids of the main patent, which are transformed or transformable by a given cloned sequence of DNA, are characterized in that they contain, on the one hand, at least a part of the genetic heritage of a line of primary cells naturally favorable to the expression of natural genes coding for a protein constituted of the polypeptide coded by the aforesaid cloned DNA sequence or containing in its structure a polypeptide sequence identical or analogous to that of this polypeptide and, on the other hand, a genetic marker permitting them to grow in a selective medium or containing an active principle normally lethal to the cells from which the hybrid is derived, but able to be inactivated by the polypeptide expressed by the said genetic marker.
The expression "primary cells" as used in the preceding designates notably any cell taken from a mammal and known for its capacity to be the seat of production of the protein or polypeptide corresponding to this protein whose production is sought. These primary cells are characterized by the fact that they scarcely multiply in vitro, that their proliferation is at most limited to the production of monolayers, the development then being interrupted by "contact inhibition" or analog. This being so, these cells can, however, in any case, be maintained alive for a certain time in appropriate mediums.
The expression "established eukaryotic cells" refers to eukaryotic cells, more particularly mammalian cells which may be cultivated in vitro and which are capable of multiplying themselves over several generations.
The process according to the invention of the parent application, Ser. No. 06/800,080, for the formation of a cellular hybrid expressing or being rendered capable of expressing a given cloned sequence of DNA was characterized: their hybridization, on the one hand, of the established eukaryotic cells, in which the aforesaid cloned DNA sequence is expressable or expressed and, on the other hand, of primary cells naturally favorable to the expression of natural genes coding for a protein constituted of a polypeptide coded by the aforesaid cloned DNA, or containing within its own structure a polypeptide sequence identical or analogous to that of this polypeptide, this co-culture being realized in a medium which does not permit the development of the said established cells, when they are not complemented by an appropriate genetic sequence, have previously been rendered able to provide the said appropriate genetic sequence to at least a part of the cell hybrids formed in the said culture medium and in that the cell hybrids formed are collected.
The invention of the parent application, Ser. No. 800,080 therefore provided modified micro-organisms transformed or transformable by cloned DNA sequences by using genetic engineering techniques, these modified micro-organisms however being such that the cloned DNA sequence in fact finds itself replaced in a genetic environment (intervening for example at the level of the differentiation or programation of the genomes containing an equivalent or identical DNA sequence) close to the natural conditions which in general prove particularly favorable to its expression.
The aim of the present invention is to provide a particularly preferred category of cellular hybrids, derived from mon

REFERENCES:
patent: 4608339 (1986-08-01), Yoakum et al.
Dubois et al., Proc. Natl. Acad. Sci., vol. 77, No. 8, pp. 4549-4553 (1980).
Houssais et al., C.R. Acad. Sci., vol. 297, pp. 497-500 (1983).
Kohler et al., Eur. J. Immunol., vol. 6, pp. 511-519 (1976).
Ochi et al., Proc. Natl. Acad. Sci., vol. 80, pp. 6351-6355 (1983).
Colbere-Garapin et al., "Late Transient Expression of Human Hepatitis B Virus Genes in Monkey Cells", EMBO J. 2(1): 21-25, 1983.
Carloni et al., "A Transformed Vero Cell Line Stably Producing the Hepatitis B Virus Surface Antigen", Gene 31: 49-57, 1984.

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