The expression of human catalytic proteins in micro algae on a commercial scale

U.S. Army Medical Research Inst Chem Def, Aberdeen Proving Ground MD

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Abstract

Bioscavenging of organophosphate (OP) by human proteins is emerging as a promising medical intervention[unreadable] for prophylaxis and post-exposure treatment against chemical warfare nerve agents. The best-.studied[unreadable] bioscavengers (BSCs) to date, meeting considerable success in pre-clinical research, are human[unreadable] cholinesterases (ChEs). However, ChEs, which are highly efficient in binding and sequestering OPs, are also[unreadable] inactivated by the toxins and therefore administration of large amounts of protein is necessary for full[unreadable] protection, raising the question of the practicality of this approach. The development of improved biocatalysts[unreadable] (BCT) (paraoxonase 1) that can catalytically degrade OPs may address this concern.[unreadable] The proposed effort offers a novel means to biomanufacture recombinant BSCs and BCTs based on the[unreadable] human proteins, butyrylcholinesterase and paraoxonase 1. In collaboration with the other projects[unreadable] participating in the Center, the capacity of these proteins to sequester or hydrolyze OPs will be improved by[unreadable] subjecting their genes to either random in vitro evolution or rational mutagenesis. To that end, we will[unreadable] undertake high-throughput screening of mutant enzyme libraries, in particular to improve the stability and[unreadable] drug-like properties of paraoxonase I. In addition, post-translational glycosylation systems or protein[unreadable] PEGylation procedures will be developed to increase the circulating lifetimes and to eliminate potential[unreadable] antigenicity of proteins produced in non-human, recombinant organisms. The OSU team (Project 6) will[unreadable] utilize the transgenic microalgae, Chlamydomonas reinhardtii, to produce pilot-plant scale quantities of these[unreadable] products for direct recovery from the culture media. In addition, they will optimize this production system for[unreadable] scale-up to commercial production capacity. The primary significance of this project is that it will develop[unreadable] pilot-plant systems for the production of enhanced, second-generation, improved ChE-based BSC and BCT[unreadable] products suitable for human clinical trials.