SBIR-STTR Award

To fully realize the benefits of genetically engineered proteins, manufacturing costs and development time must be significantly reduced. Technologies that combine robust expression, and inexpensive methods of purification are key to this goal. This Phase I proposal will demonstrate the feasibility of Tetrahymena thermophila, a common pondwater ciliate, as a novel system for low cost production of recombinant proteins using strategies that focus on a "model" vaccine antigen with considerable importance in human and animal health, namely, the hemagglutinin protein (HA) of influenza virus. Our specific goals are to 1) introduce the HA gene from the current H5N1 strain of avian flu into T. thermophilia 2) screen recombinant cell lines for the expression of HA using antibodies specific for the protein; and 3) devise rapid, inexpensive methods for downstream purification of candidate vaccine antigens.

Keywords:
TETRAHYMENA, RECOMBINANT PROTEIN EXPRESSION, HEMAGGLUTININ (HA), H5N1, COST-EFFECTIVE, HIGH-YIELD

Recombinant proteins are critically important in a wide range of applications that extend from the treatment of human and animal disease, to chemical and biological defense. Production of genetically engineered vaccine antigens, therapeutic proteins (including monoclonal antibodies), industrial enzymes, biopolymers, and bioremediation agents now constitutes a multibillion dollar-per-year industry. However, the need for low cost systems for the rapid production of these proteins remains great. In Phase I we demonstrated feasibility of the use of a free-living ciliate, Tetrahymena thermophila, as novel platform for high-level expression of the H5 hemagglutinin of avian influenza virus. In Phase II we will extend this work to commercially relevant enzymes, and antibody fragments targeted against biowarfare agents with the goal of benchmarking both yield and production costs against competing systems. In addition, we will develop user-friendly protein expression kits for the research tool market based on the TetraExpressTM platform.

Keywords:
Tetrahymena, Protein Expression, Rapid, Cost-Effective, High-Yield, Glycosylation