Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1997-09-25
2000-10-24
Scheiner, Laurie
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 697, 435 698, 4242181, C12N 1509
Patent
active
061365610
ABSTRACT:
The Flaviviridae comprise a number of medically important pathogens that cause significant morbidity in humans including the dengue (DEN) virus, Japanese encephalitis (JE) virus, tick-borne encephalitis virus (TBE), and yellow fever virus (YF). Flaviviruses are generally transmitted to vertebrates by chronically infected mosquito or tick vectors. The viral particle which is enveloped by host cell membranes, comprises a single positive strand genomic RNA and the structural capsid (CA), membrane (M), and envelope (E) proteins. The E and M proteins are found on the surface of the virion where they are anchored in the membrane. Mature E is glycosylated and contains functional domains responsible for cell surface attachment and intraendosomal fusion activities. Problems have arisen in the art with respect to producing recombinant forms of the E glycoprotein that retain their native configuration and attendant properties associated therewith (i.e., ability to induce neutralizing antibody responses). To date, recombinantly produced E glycoproteins have suffered from a number of limitations including improper glycosylation, folding, and disulfide bond formation. The claimed invention has addressed these concerns by providing secreted recombinant forms of the E glycoprotein that are highly immunogenic and appear to retain their native configuration. Carboxy-terminally truncated forms of E containing the amino terminal 395 amino acids and a suitable secretion signal sequence were generated in Drosophila melanogaster Schneider cell lines. The recombinant proteins produced by this expression system should prove useful, inter alia, as immunogens and diagnostic reagents.
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Clements David
Ivy John
Nakano Eilen
Hawaii Biotechnology Group, Inc.
Parkin Jeffrey S.
Scheiner Laurie
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