Abstract: The present invention discloses and claims vaccines containing, as an active ingredient, a secreted recombinantly produced dimeric form of truncated flaviviral envelope protein. The vaccines are capable of eliciting the production of neutralizing antibodies against flaviviruses. The dimeric forms of truncated flaviviral envelope protein are formed 1) by directly linking two tandem copies of 80% E in a head to tail fashion via a flexible tether; 2) via the formation of a leucine zipper domain through the homodimeric association of two leucine zipper helices each fused to the carboxy terminus of an 80% E molecule; or 3) via the formation of a non-covalently associated four-helix bundle domain formed upon association of two helix-turn-helix moieties each attached to the carboxy terminus of an 80% E molecule.

Abstract: An assay is disclosed for determining whether a test compound modulates the activity of an enzyme having a metallated active site. The assay method employs a comparison of the binding ability of the metallated and unmetallated forms of the enzyme to the test compound.

Abstract: The recombinant expression and secretion from eucaryotic host cells, particularly Drosophila cells, of Flavivirus nonstructural (NS) protein, particularly NS1, is useful in combination with Flavivirus truncated envelope (E) protein to protect a host subject from infection and disease from Flavivirus species. Further, NS1 is useful as a diagnostic of flaviviral infection. Compositions of truncated flaviviral envelope protein and flaviviral nonstructural protein induce high titer virus neutralizing antibodies believed to be important in protection against flaviviral infection and which are useful in diagnosis of infection by the virus.

Abstract: A recombinant DNA encoding coffee bean &agr;-galactosidase permits the production of purified forms of this protein. The protein is useful in converting human Type B red blood cells into cells physiologically similar to Type O red blood cells. The availability of this enzyme permits more effective conversion than use of &agr;-galactosidase from other sources.

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).

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).

Abstract: Neurospora crassa transformations are employed for production of heterologous proteins, where DNA constructs are prepared using a foreign gene under transcriptional and translational regulatory regions functional in Neurospora. A functional signal sequence is provided for secretion of the heterologous gene product. An auxotrophic Neurospora host is employed and transformed by the combination of the subject construct and a complementing gene. The resulting transformants provide for efficient secretion of the heterologous product.

Abstract: Immunoaffinity purifications are performed using non-aqueous media, desirably, a small amount of an aqueous buffered medium, and, optionally, a surfactant. Hydrophobic ligands can be extracted from samples, immunoaffinity purified and eluted to provide a substantially pure product.

Abstract: Novel specific protein binding assays are provided employing a solvent system comprising a solvent characterized as hydrophobic, essentially anhydrous and water immiscible. Particularly, lipophilic analytes are shown to be sensitively determined using antibodies in hydrocarbon media, optionally substantially saturated with aqueous buffer and/or comprising an anionic surfactant.