As part of a CRO, I am involved in the development of the following methods:Size Exclusion ChromatographyHydrophobic Interaction ChromatographyIon Exchange ChromatographyReversed Phase ChromatographyCapillary Electrophoresis (SDS and IEF) IEF and SDS-PAGE gelsBCAA280GCDLSHACH TurbidimeterProtein Formulation Optimization

Evaluation of protein quality for Process Development:- Capillary Electrophoresis (CE-SDS)- Size Exclusion Chromatography (SEC)- Glycan Analysis by 2-AB labeling and HPLC- Circular Dichroism (CD)- Differential Scanning Calorimetry (DSC)

- Protein HPLC Method Development: Anion and Cation Exchange, SEC, Reversed Phase.(Shimadzu Prominence)- Protein Reduction/Alkylation- SDS Page- Aseptic Handling- Endotoxin Evaluation and Removal- Starch assay- GMP practices and evaluating SOPs

Small Molecule Characterization• Evaluated methods for determining compound aggregation (Wyatt DynaPro DLS and Corning EPIC). Tested 1400 compounds and compared reults as a factor of compound class and pH.• Solubility determination of organic compounds using nephelometry. Protein-Ligand Interactions• Determination of Kd's for protein ligand binding by Isothermal Titration Calorimetry (ITC) and thermal shift assays (DSF and CD). • Optimized an HPLC/MS method to monitor inhibitors of PCSK9 enzymatic self cleavage (cholesterol program).• Developed data analysis programs for Kd and enzymatic analysis.Protein Characterization• Responsible for the operation of Perkin-Elmer 494 N-terminal protein sequencer.• Experience in HPLC-MS (Agilent 1100 HPLC with Waters Micromass LCT).

Six patents awarded in protein drug delivery, formulations, and small molecules used against protein targets.Small Molecule Characterization• Initiated Schering-Plough’s high-throughput solubility method for organic compounds using nephelometry. Evaluated solubility results with respect to pK, logP, and logD predictions. Developed an automated method for determining pH dependent solubility.• Initiated program for determination of LogP using the Sirius GlpKa instrument.• Patent approved for everninomicin formulations, using light-scattering to determine conditions to obtain a monomeric form of the compound.Protein-Ligand Interactions• Determination of Kd's for protein ligand binding by Isothermal Titration Calorimetry (ITC) and thermal shift assays (DSF and CD). Two patents.• Developed a novel thermal shift assay for the evaluation of covalent inhibitors of HCV NS5B polymerase, critical for improving SAR in providing a recommendation for development. • Optimized an HPLC/MS method to monitor inhibitors of PCSK9 enzymatic self cleavage (cholesterol program). Improved patented method.• Developed data analysis programs for Kd and enzymatic analysis.Protein Characterization• Responsible for the operation of Perkin-Elmer 494 N-terminal protein sequencer.• Experience in HPLC-MS (Agilent 1100 HPLC with Waters Micromass LCT).• Provided analytical support using the following protein analysis techniques: IEF, SDS-PAGE, ELISA, electroblotting, protein digestion, MALDI-TOF.Drug Delivery• Patent approved for the attachment of polyethylene glycol to interferon for use as a sustained release drug delivery system.• Two patents approved for sustained release drug delivery of crystalline interferon.• Developed a procedure for preparing microcapsules containing interferon as an injectable drug delivery system. Resulted in peak levels of interferon in monkeys at three days versus 24 hours for unencapsulated.