1. Biodefense Vaccines, Therapeutics and Diagnostics NGIC Workshop September 27, 2004 Robert L. McKown, Ph.D.

2. What is Biotechnology?

3. Recombinant genetic Engineering…using biological process to develop products G. Steven Burrill 1997 Genetic Engineering Raw Materials Living System Product

4. Enabling Technology Recombinant DNA Technology Boyer and Cohen, 1973

5. The Birth of an Industry University of California, San Francisco Genentech Inc. South San Francisco

6. Founded April 7, 1976 Herbert Boyer and Robert Swanson

7. Proof of Concept Cloning the human insulin gene Cloned by Genentech scientists in 1978 Licensed to Eli Lilly First recombinant drug marketed, 1982 Product revenues from human insulin (humulin) reached $900 million in 2003

8. The Promise of Biotechnology Bioproduction of molecules so complex they can only be synthesized in a living system DNAPROTEIN

9. Biotech Products

10. The JMU Biomanufacturing Laboratory Concept document created in 1997 Development funding awarded in 1998 Lab space committed in 1999 Lab design and set up in 2000 Became operational in 2001

11. Founding Sponsors

12. Biomanufacturing Technology Genetic manipulations and analysis Cell line development Fermentation and cell culture technology Product purification technology Quality assurance and quality control Regulatory guidelines and compliance Process development

13. The Production Platform Cell Line Development Systems Management Regulatory Affairs Quality Assurance Production Product Recovery Product Analysis Quality Control Analytical Testing Documentation

14. Development of Expression Systems

15. Cell Line Development

16. Production and Purification

17. Product Analysis

18. Proof of Concept Cloning, Expression, & Purification of Green Fluorescence Protein (GFP)

19. Purification of GFP

20. Analysis of GFP Fraction I II III IV STD Mol. Wt. 94,000 67,000 43,000 30,000 20,100 14,400

21. Projects and Collaborations Lacritin project, UVA and EyeRx Vaccine and diagnostics projects, USAMRIID Staphylococcus aureus Protein A project, Genentech, Inc. New River Pharmaceuticals project M. tuberculosis project, Veterans Hospital, Long Beach, CA

22. Research and Development of a Recombinant Protein (Lacritin) as a New Therapeutic for Treatment for Dry Eye Syndrome

23. Lacritin and the Lacrimal Gland A stable, novel human tear glycoprotein Produced primarily by the lacrimal gland located on the upper eyelid Also found in salivary glands and the thyroid

24. Properties of Lacritin Demonstrated in Cell Culture Lacritin hydrates the corneal surface as a natural component of tears Lacritin causes the rapid, sustained release of calcium in cultured corneal epithelial cells Lacritin stimulates proliferation of ductal cells

25. Potential as a Therapeutic Agent Dry Eye Syndrome Caused by the decrease in both the quality and volume of tear secretion Affects over 35 million Americans The application of eye drops is the primary current treatment Lacritin as a Potential Therapeutic Promotion of tear production Induction of cell proliferation

26. Research History Lacritin gene discovered and characterized by the Gordon Laurie lab at UVA JMU collaboration with the Laurie lab Engineered a recombinant mature lacritin gene that was cloned, expressed, and purified Current Research Creation of C-terminal deletions to identify domains of biological activity Lacritin protein in pre-clinical animal studies for treatment of Dry Eye Syndrome

27. Future Applications of Biotechnology New Human Therapeutics and Vaccines New Human Diagnostics New Genetically Engineered Microbes, Plants, and Animals New Biomaterials (Rubber, Plastics, Spider Silk) Genetically Engineered Humans?

28. Acknowledgements Robert McKown, Ph.D., JMU Ron Raab, Ph.D., JMU George Coffman, Ph.D., JMU Gordon Laurie, Ph.D., UVA Jeremy Goodin, Ph.D., USAMRIID Patricia Williams, Ph.D., EyeRx JMU Students