Application development foci; Very briefly new product development and application development focused primarily on LCMS and also on LCUV of small and mostly large molecules, such as biologics including but not limited to mAbs and oligos and effective use of consumables.

Hands-on team leader: LCMS analyses of small and especially large molecules to satisfiy regulatory requirements and SOPs in a GLP environment. Creator and developer of a LC-HRAMS service line, focused on small molecule and especially protein bioanalyses.Accomplishments• Building and leading a new laboratory team• Method development, qualification, and application• Creating quotes, conversion to contracts • Preparing scientific marketing presentations• Training and mentoring of colleagues on standards preparation and documentation• Leader for laboratory safety and cleanliness• LC-MS analysis of pharmaceuticals, including PK, PD, metabolite ID / structure, stability, anti-drug antibody measurement, protein activity assays and more• Identifying and quantifying sites of covalent conjugation of compounds to proteins, and number of tagged sites per protein molecule• Operation, maintenance and troubleshooting of LCMS instrumentation• Use of Tomtec Quadras, evaporators, analytical balances, shakers, evaporators and more• Optimization of MRM transitions, PRM methods, and signature peptide analyses• Optimization of LC methods • Development of extraction methods from biological matrices • Writing and developing bioanalytical procedures and SOPs • Mastery of key software

Development and application of discovery proteomics, LCMS-biologicals analyses and quantitative bioanalytical LCMS based analyses of small and large molecules, and other applicable laboratory procedures are my central foci. This collaborative work is guided by, and contributes to basic biomedical discoveries, regulatory requirements and company standard operating procedures to satisfy good laboratory practice (GLP) standards. I rigorously and clearly document all of my work, also in accordance with GLP.Accomplishments• As the leader of projects, creating and maintaining client satisfaction• Establishment and lead on new LCMS-based protein analysis services; intact and bottom-up using UHPLC-MS/MS and high-resolution/accurate mass instrumentation• LC-MS/MS analysis of many pharmaceuticals, including sample preparation, analysis and quantification• Effective use of HPLC, UHPLC triple quadrupole and Orbitrap LCMS technologies• Rapid, collaborative, efficient method development, including effective and punctual troubleshooting
• Superb performance of and improvements to analyte extraction protocols from a wide variety of biological matrices• Preparation, labeling, tracking, reviewing, storing, retrieving and analyzing reference standards, GLP-compliant reagents, control matrices, and quality controls• Training and mentoring of colleagues on the application of established and newly developed methods, providing technical advice and troubleshooting• Review of the work of colleagues, also according to GLP standards• Contributing to communication between our laboratory and sponsors (clients)• Creating, evaluating, refining and clarifying bioanalytical protocols• Understanding, providing leadership on, and complying with all applicable laboratory safety and cleanliness procedures and policies

• Leading dozens of proteomics projects, providing consultation, creating experimental plans, maximizing sample quality and biological relevance, generating, analyzing interpreting and reporting data• Developing and applying many effective, robust, quantitative LC-MS/MS methods, including multidimensional LC- (MDLC) MS/MS for proteomic analyses• Improving label-free, iTRAQ and SILAC quantification and statistical analyses of protein abundance• Creating a unique chemistry enabling identification and quantification of many proteins which otherwise eluded detection• Specifying custom programming objectives; overseeing development of data analysis scripts and a new website; identifying improvements and evaluating the results• Publishing articles in peer-reviewed journals; full Professor-level writing and figure preparation skills• Training and supervising technicians, graduate students, postdoctoral fellows and staff scientists; contributing to and overseeing their method development efforts• Developing relationships and collaborations with vendors; obtaining 22%-60% discounts• Developing budgets• Creating position descriptions, hiring scientific personnel• Preparing grant applications; providing letters of support• Reviewing manuscripts and grant applications• Delivering lectures; leading scientific discussions

• Leading dozens of proteomics projects, providing consultation, creating experimental plans, maximizing sample quality and biological relevance, generating, analyzing interpreting and reporting data• Developing and applying many effective, robust, quantitative LC-MS/MS methods, including multidimensional LC- (MDLC) MS/MS for proteomic analyses• Improving label-free, iTRAQ and SILAC quantification and statistical analyses of protein abundance• Creating a unique chemistry enabling identification and quantification of many proteins which otherwise eluded detection• Specifying custom programming objectives; overseeing development of data analysis scripts and a new website; identifying improvements and evaluating the results• Publishing articles in peer-reviewed journals; full Professor-level writing and figure preparation skills• Training and supervising technicians, graduate students, postdoctoral fellows and staff scientists; contributing to and overseeing their method development efforts• Acquiring, managing installation and service, as well as operating MDLC-MS/MS instrumentation• Developing relationships and collaborations with vendors; obtaining 22%-60% discounts• Developing budgets• Creating position descriptions, hiring scientific personnel• Preparing grant applications; providing letters of support• Reviewing manuscripts and grant applications• Delivering lectures; leading scientific discussions

Relevance and summaryDynamic protein phosphorylation and de-phosphorylation are critical to cell signaling. Among many projects, we developed and applied some of the early MDLC-MS/MS tools for analysis of phosphoproteins. We also developed and applied additional proteomic methods.Accomplishments• Co-directed and participated in the identification of phosphorylated proteins that, in combination with pathway analyses, successfully predicted novel culture conditions facilitating hESC self renewal.• Identified and quantified phosphorylation sites on thousands of proteins, and determined differences in protein phosphorylation events between different cell states, cell lines and signaling pathways• Managed and performed collaborations that improved MDLC-MS/MS• Led efforts to validate MS/MS spectra from cancer and embryonic stem cell lines. Hired two interns to assist; trained and supervised four people in this work• Supervised one summer intern, one research associate, five postdoctoral fellows and one staff scientist in culturing, treating, and preparing proteins from mammalian cells• Initiated and performed experiments for, and supervised a postdoctoral fellow on, a project to investigate Insulin-like growth factor 1 signaling in cancer cells, and directed the nexus of the proteomic and cell biological studies• Initiated hydrogen/deuterium exchange MS/MS analyses to support structural biology

Relevance and summaryKnowledge of the structures of membrane proteins has been especially limited. My research consisted of biochemical and biophysical studies of a model membrane protein, the Tobacco Mosaic Virus (TMV) Movement Protein (MP), which is essential for cell-cell spread of infection. Accomplishments• Expressed recombinant TMV MP in E. coli, improved the yield ca. 10-fold• Established and further developed a protocol for MP purification, including anion exchange chromatography and preparative SDS-PAGE; demonstrated MP purity and monodispersity using gel filtration chromatography, analytical ultracentrifugation and electron microscopy• Supervised a research assistant; published the work in a peer-reviewed journal• Formulated and refined the first topological model of any MP using hydropathy analysis, CD spectroscopy, partial proteolysis, western blotting, MALDI-TOF MS and analytical ultracentrifugation

Relevance and summaryLectin genes encode carbohydrate-binding proteins. In spite of extensive studies, their biological function(s) are often unclear. My research included cloning, characterizing and testing functions of lectin genes in Medicago sativa (alfalfa). The M. sativa lectin genes were found to have important, unanticipated functions in symbiosis, development, reproduction, and defense.Accomplishments• Initiated, performed, and collaborated with others on a new lectin molecular biology project:o Constructed and screened DNA librarieso Cloned and characterized lectin geneso Constructed recombinant DNA plasmidso Explored lectin gene functions using transgenic plantso Demonstrated variable expression of lectin mRNAs in transgenic plants but decreased accumulation of a putative lectin protein in antisense plants, which correlated with abnormal phenotypes• Increased the yield of transgenic plants ca. 5-fold.

I devised, initiated, developed and successfully implemented quantitative immunochemical diagnostics using antibodies that I generated and ELISA that I developed.

As a graduate research assistant, I was responsible for creation of, performance of, and publishing a new quantitative immunochemical diagnostics project.