Advanced Bionutrition Corporation

Martek Biosciences Corporation

F C Thomas Allnutt

David J Kyle

Kirk Emil Apt

Arun Dhar

Ruth Barratt

Robert Bullis

James Casey Lippmeier

John Peter Morseman

Robert M Bowers

Mark Wesley Moss

James Casy Lippmeier

Infectious pancreatic necrosis virus (IPNV), the etiologic agent of infectious pancreatic necrosis in salmonid fish, causes significant losses to the aquaculture industry. The gene for the viral capsid protein (VP2) was cloned into a yeast expression vector and expressed in. Expression of the capsid gene in yeast resulted in formation of approximately 20 nanometer sub-viral particles composed solely of VP2 protein. Anti-IPNV antibodies were detected in rainbow trout vaccinated either by injection of purified VP2-subviral particles (rVP2-SVP) or by feeding recombinant yeast expressing rVP2-SVP. Challenge of rVP2-SVP immunized trout with a heterologous IPNV strain and subsequent viral load determination showed that both injection and orally vaccinated fish had lower IPNV loads than naive or sham-vaccinated fish. This study demonstrates the ability of rVP2-SVPs to induce a specific immune response and the ability of immunized fish to reduce the viral load after an experimentally induced IPNV infection. The invention is not limited to IPNV, and is applicable to other similar viruses for which SVPs can be made and administered to fish.

Antigenicity of infectious pancreatic necrosis virus VP2 sub-viral particles expressed in yeast

Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically-derived energy. In this study it is shown that the microalgacan be engineered to import glucose and grow in the dark through the introduction of genes encoding glucose transporters. Both the human andglucose transporters facilitated the uptake of glucose by, allowing the cells to metabolize exogenous organic carbon and thrive, independent of light. This is the first successful trophic conversion of an obligate photoautotroph through metabolic engineering, and it demonstrates that methods of cell nourishment can be fundamentally altered with the introduction of a single gene. Since strains transformed with the glucose transport genes are able to grow non-photosynthetically, they can be exploited for the analysis of photosynthetic processes through mutant generation and characterization. Finally, this work also represents critical progress toward large-scale commercial exploitation of obligate phototrophic algae through the use of microbial fermentation technology, eliminating significant limitations resulting from light-dependent growth.

Trophic conversion of obligate phototrophic algae through metabolic engineering

Methods and constructs for genetic manipulation of one or more of shrimp, shellfish, mollusks, and fish are disclosed. The nucleic acid construct includes a promoter and an internal ribosome entry site of an insect picomavirus, such as a cricket paralysis-like picomavirus. One or more open reading frames can be operably associated with one or both of the promoter and the internal ribosome entry site, and one or more proteins or protein subunits can be expressed upon introduction of the construct into a host cell, such as into a shrimp. Method for producing immortalized crustacean cell lines using enhancer elements derived from shrimp and/or shrimp viruses are also described.

Crustacean expression vector

A crustacean or rotifer is infected with a recombinant infectious virus that expresses a protein exogenous to the virus. The genome of the crustacean or rotifer itself remains unaltered. Crustacean, rotifer, insect, or viral promoters drive the transcription of a gene inserted into the recombinant virus genome, and the virus replicates in the crustacean or rotifer cell cytoplasm. The infected crustacean or rotifer can be provided directly to humans or non-human animals, or, following production and harvest of the crustaceans or rotifers, purified recombinant protein or polypeptide can be provided. Large quantities of biopharmaceuticals can be produced rapidly and inexpensively using this production system.

Crustaceans as production systems for therapeutic proteins

Transfected shrimp as production systems for therapeutic proteins

In a method for quantitating an analyte by measuring time resolved transfer of fluorescence energy to or from a label quantitatively associated with the analyte, the present invention provides an improvement comprising measuring the energy transferred from donor compounds having the ability to absorb light energy and then transfer this energy to cross-linked allophycocyanin in a time-resolved manner, where the cross-linked allophycocyanin used according to this invention has not been exposed to strongly chaotropic agents after cross-linking.

Growing community of inventors

Glenwood, MD, United States of America

F C Thomas Allnutt