SBIR-STTR Award

This Phase I Small Business Innovative Research project will involve the construction of DNA expression vectors that will facilitate the production and secretion of high levels of peptides and other fermentation products in bacteria. The vectors will allow for simultaneous production and secretion of the target peptide and a reporter peptide. A hallmark of the proposed project is the development of an on-line, real-time, fermentation monitoring system that will allow improved process control during the fermentation process. The proposed research will address the need for improved large-scale production of a wide range of peptides, including antibodies for detection and defense of chemical and biological weapons, and industrial enzymes for biocatalysis. The feasibility of this approach will be demonstrated by constructing the expression vectors and producing a commercially important lipase enzyme that can be used for creating enantio-pure feedstocks for the production of pharmaceutical and fine chemicals. The products of this Phase I research will be a series of versatile cassette-based expression vectors and host organisms designed for optimum expression and secretion of peptides, and a design for an on-line monitoring and process control system for industrial-scale fermentation. Both the vectors and fermentation control systems developed during this project will have a wide range of government and private sector applications. The technology will be useful for large-scale production of antibodies for detecting and inactivating chemical and biological warfare agents, and it will be useful for producing commercial quantities of a wide range industrial enzymes and non-proteinacious fermentation products.

The goal of this Phase II SBIR project is to develop an improved process monitoring and control system for biomanufacturing. The completed system will include expression vectors that can be used to produce and secrete commercially useful peptides and other fermentation products, and a process control system for large-scale biomanufacturing. The vectors will encode for the simultaneous expression and secretion of a target peptide and a reporter peptide that can be monitored on-line during fermentation to facilitate process control. The process control system will incorporate an enzyme assay and detector module to monitor the production of the target peptides and to control the fermentation process. The vectors and process control system will be used to produce antibodies and peptides for a wide range of commercial and defense applications. During Phase II, we will complete construction of Gram negative, Gram positive, and broad-host-range expression vectors and couple them to suitable host organisms. We also will complete development of the fermentation control module, and perform pilot-scale fermentation experiments to calibrate the process monitoring system and to produce fermentation products in quantities sufficient to evaluation their commercial and defense utility

Keywords:
BIOMANUFACTURING, PROCESS CONTROL, BIOCATALYSIS, ANTIBODIES, FERMENTATION, EXPRESSION VECTORS