Life Sciences. Professional Information. Resume. Creative Problem-Solver, Challenge Enthusiast. Member Number: West Bloomfield, MI 48324

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1 Career Portfolio provided by Beyond.com Life Sciences Creative Problem-Solver, Challenge Enthusiast West Bloomfield, MI Member Number: Portfolio: Professional Information Job Function: Science & Biotech, Research Education: Post Graduate Degree Experience: 5 10 Years Employment: Full-Time Salary: $70-150k Security Clearance: None Citizenship: U.S. Citizen Resume CAREER OBJECTIVE To aid a risk-taking company in providing innovative solutions utilizing my melded background in chemical engineering, chemistry and molecular biology established during my academic laboratory and industrial career. EDUCATION Orchard Lake St. Mary s Preparatory Orchard Lake Village, Michigan 4/*** University of Michigan BSChE 1/*** Ann Arbor, Michigan Chemical Engineering Wayne State University PhD 5/*** Molecular Biology, minor in Chemistry GPA: *** 5/*** Graduate Certificate in Business Administration GPA: ***

2 LABORATORY EXPERIENCE Wayne State University, Dr. Philip Cunningham Laboratory Post-Doctoral Researcher *** to present Commercialization of antimicrobial peptides and their method of isolation outlined in the research project below. Graduate Research Assistant *** to *** Outstanding Research Assistant Award, Biological Sciences, WSU*** ***, *** Research Projects *** In vivo display: An irrational approach to discover de novo antimicrobials. I developed a novel in vivo drug-discovery technology that bypasses the previous issues of membrane permeability, pre-screen target identification and physiological molecular interactions by screening *** s of millions of peptides in vivo for antimicrobial effects on Escherichia coli. I produced a library of plasmids from expression in E. coli encoding a display protein attached to a random peptide. Via high-throughput screening in liquid media the method yielded peptides whose expression is detrimental to bacterial growth in one of three phenotypes: bactericidal, bacteriostatic and bacteriolytic. Further analysis of enriched peptides for target binding, peptide sequence, minimum inhibitory concentrations, induced growth curves and spectrum efficacy characterized the potential of each peptide to serve as a new lead for antimicrobial development. Using this new method I have discovered hundreds of peptides with antimicrobial activity against E. coli. Three such peptides were studied against other organisms as drug-like agents and found to have activity against Gram-negative organisms such as Shigella sonnei and Salmonella typhimurium and Gram-positive organisms such as Staphylococcus aureus, Enterococcus faecalis and Streptococcus salivarius. None of the three peptides demonstrated hemolytic activity making them great leads for the development of new antimicrobials. I also adapted the screen for use in finding antimicrobial peptides in the periplasm, nutrient starvation, anaerobic conditions and in the reverse orientation on the display protein (N versus C-terminus). The latter derivative required the innovation of using in vivo TEV protease expression to cleave off a protective peptide N-terminal to the peptide library to negate translation efficiency problems with randomizing the 5 of the encoding mrna. This powerful screen can be adapted for use in other organisms such

3 as S. aureus to isolate species specific or narrow spectrum antimicrobials. I utilized MS/MS to identify targets bound to antimicrobial peptides, affinity columns (step, histidine tags) to purify peptide-target complexes, synthetic peptides for MIC s, mutational adaptation to antimicrobial and mechanism of action. *** Investigation of a central domain pseudoknot in the ***S ribosomal subunit. Project objective was to determine the functional importance of the base pairing regions of a particular pseudoknot in the central domain of the ***S in E. coli. A total of *** mutations were created by site-directed mutagenesis and a succession of cloning reactions. Assays to detect function were performed on each mutant to analyze mutation complementation and thermodynamic interactions. The structure of the mutant ribosomes were analyzed using homology modeling software and assembly/association defects were identified via ribosome sucrose gradients. *** Isolation of non-functional ***S rrna single mutants. The ***S ribosomal subunit has served as a reliable antimicrobial target for the discovery and development of new antimicrobials. A follow-up to the Cunningham Laboratory s instant evolution system was used to elucidate new drug targets in the bacterial ***S ribosomal subunit resulting in the discovery of single mutations that completely negated function of the ribosome in vivo. These mutations, when compared to those in the human ***S ribosomal subunit, pinpoint new potential drug targets for the development of de novo antimicrobials. Equipment Beckman Coulter robotics (Biomek FX), Gene Machines Higro ***well plate incubator, Licor ***L DNA sequencer, fluorometer and spectrophotometer microplate readers, thermocyclers, SLM french press, Sorvall RC***C high speed centrifuge, Sorvall Wf ultrafuge, GE Typhoon FLA *** fluorescent microscope. Methods Circular Dichroism, peptide synthesis, minimum inhibitory concentrations, fluorescent microscopy Polymerase Chain Reaction, site-directed mutagenesis, DNA restriction digests, cloning, subcloning, alkaline lysis plasmid preparation, DNA isolation using gel extraction protocols, Sanger sequencing reactions, sequence analysis, reporter gene assays, electroporation, electro-competent cell preparation, PFU polymerase preparation, Acrylamide gel and reagent preparation, SDS PAGE gels, ribosome preparation, antibiotic preparation, M*** phage display and library expansion.

4 Software Primer design software (Oligo DT), Genetic cloning and sequence software (Gene Construction Kit), Molecular structure software (PyMol, PDB Swiss viewer), DNA sequencing analysis software (Licore, Align IR, e-seq), peptide modeling, motif searching Microsoft Excel, PowerPoint, Word. WORK EXPERIENCE Wayne State University Staff*** Graduate Teaching Assistant****** to present ***oductory microbiology lab instructor of *** undergraduate students for 6 semesters*** Prepared weekly lectures, discussions, quizzes, exams and posted and recorded grades. Learned to work with and educate students who had minimal prior knowledge of biology. ***nced molecular biology lab instructor of *** graduate for 6 semesters Discussed and oversaw advanced experiments and methods conducted by graduate level students. Assisted in the design and supply of reagents necessary for advanced experiments and methods. Wrote, designed and delivered lectures/experiments in the absence of course s professor. Graduate Research Assistant: Dr. Robert Arking Laboratory*** *** to *** I conducted experiments on D. melanogaster demonstrating the linkage of aging to radical superoxides in mitochondria. I changed food, identified mutants, measured life spans, constructed databases of data and handled paraquat. DNA Software Inc.*** Wet Laboratory Technician*** *** *** to *** I developed and confirmed a system for identifying single mutations in the ***S ribosome that negated function (See Project 3 above). The data used to confirm client s RNA structure prediction program. Mainline Technology Inc., IM Diagnostics Inc.*** Lab Supervisor****** to present Ann Arbor, Michigan I developed and wrote SOP s and guidelines for the formulation of reagents, buffers and control solutions distributed with molecular diagnostic kits. I carried out the production of reagents, buffers and control solutions required for commercial molecular diagnostic kits for pregnancy (hcg), streptococcal (Protein A), drugs of abuse and Epstein-Barr. I grew, harvested and prepared cultures of Streptococcus pyogenes and S. agalactiae for use in control solutions. I produced solutions of human chorionic gonadotropin (hcg) and luteinizing hormone (LH) for the use as control solutions and for quality analysis of molecular diagnostic kits via ELISA. I setup small independent research and development lab focused on molecular diagnostic and molecular biology research carrying out projects of my own design without supervision. University of Michigan*** Ann Arbor, Michigan Lab Attendant*** *** to *** I maintained food, water, and cages and followed procedures on how to euthanize and handle laboratory mice.

5 I observed and discussed experiments conducted by PhD students. PRESENTATIONS/CONFERENCES The Challenge of Antibacterial Drug Development San Diego, California American Association for Clinical Chemistry (AACC)*** *** *** Orlando, Florida Biological Sciences Department Research Retreat, WSU*** Oral and Poster Presentation (Best Poster Award)*** *** Grosse Pointe, Michigan Wayne State University Graduate Exhibition*** Biological Sciences Poster Day, WSU*** Poster Presentation (Best Poster Award)*** ***, *** RNA Rustbelt Meeting*** ***, ***, *** Mt. Sterling, Ohio RNA Club, WSU*** Oral Presentation*** ***, ***, ***, ***, *** PATENTS Mainline Technology, Inc.*** Ann Arbor, Michigan Prostate Cancer Molecular Diagnostic*** *** Unfiled due to lack of funding Wayne State University*** In vivo display system and antimicrobial peptides *** *** derived from the system (See Research Project 1 above) Pending HOBBIES

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