Protein Purification and characterization via HPLC, CE, etc.

Introduced and developed HPLC/UPLC assays for Monoclonal Antibodies (mAb) for both Cell Free synthesis and CHO based expression, created novel HPLC assays for various enzymes related to protein synthesis. Designed and implemented protein purification schemes for various vaccine proteins (EBVgb, Influenza, etc (MedImmune). Developed HPLC titer assay for EBVgb. Characterized mAb's via HPLC, UPLC, LC-MS in both Research and GMP mode.

Responsible for reviewing data packages from CMO for recombinant drug (Andexanet Alfa), issuing COAs for release of Drug Products and Drug Substances, requesting amendments to COA's from CMO's.reviewing Batch Records., communicating and requesting corrections and data. Interfacing with QA. Reviewing Stability data, issuing COA's.

Analytical Development- IgG's, Antibody Drug Conjugates: HPLC primarily. Expression is cell free extract based. Devised several new HPLC methods to quantitate various proteins involved in the cell free synthesis-some novel.

Employed as a contractor: Set up protein quality assessment capability through developing/creating protein related HPLC methods for determining quantity and quality of recombinant proteins expressed in CHO cells- without prior purification. Successfully created accurate, precise method for large (300 Kda +) heavily glycosylated protein in media. Other mammalian cell culture, transfection, expression and purification

Developing and implementing protein purification schemes for various vaccine related proteins (HA, EBVgb, etc), analyzing vaccine related proteins via, HPLC (developed methods), PAGE-SDS (electrophoresis), Western blotting. Cell Culture and Protein Expression.

Responsible for HPLC analysis of cell harvests and column fractions for mammalian and bacterial IgG and Fab's. Developed test method for determining Fab titers from bacterial broths and column fractions including HPLC column, conditions and pre-analysis sample treatment (Endonuclease digest, etc) . Responsible for extended characterization for the various IgG's produced at Xoma including intact mass determination. Developed/introduced glycan profiling and antibody mapping techniques utilizing MALDI-TOF as ancillary methodologies for CE and HPLC-MS. Developed methodologies to characterize hinge region degradation pathway. Coordinated dialysis, and outside contractor AAA analysis. Have run titer analysis (IgG and Fab) in GMP mode as well as R&D mode.

Analytical Development: Responsible for HPLC Assay Development and analysis of cell harvests and (purification) column fractions for mammalian and bacterial Monoclonal Antibodies and Fab's. Developed test methods for determining Fab titers from bacterial broths and column fractions including HPLC column, conditions and pre-analysis sample treatment Responsible for the characterization of numerous Monoclonal Antibodies produced at Xoma (charge isoforms, post-translational modifications). Developed/introduced glycan (glycosylation) profiling and antibody peptide mapping techniques utilizing MALDI-TOF as ancillary methodologies for CE and HPLC-MS. Developed HPLC and Mass Spec methodologies to characterize hinge region degradation pathway. Coordinated dialysis, and outside contractor AAA analysis. Monoclonal Antibody Analytical development and qualification for GMP use in support of Phase I and II Clinical Studies

Developing and running SEC, Reverse Phase, Ion Exchange, and Protein A HPLC, LC-MS, and MALDI-MS for various mAb's in R&D, developing assays to transfer to QC. Utilizing HPLC, MALDI-MS, and LC-MS to identify various Post Translational Modifications.

Characterization of proteins through HPLC Mass Spectrometry, MALDI-TOF Mass spec. and PAGE-Gel Electrophoresis.

Designed (in collaboration with Director) and conducted experiments in the development of nanobead -antibody protein conjugation chemistry for cytokine identification and localization (Intracellular and Cell Surface Staining projects).Evaluated several combinations of modified styrene surfaces and modified target functional groups for direct and indirect coupling after oxidative or reductive chemistry. Conventional and bead ELISA development.Testing of proprietary software developed for digital imaging and quantification of multihued nanoparticlesEvaluated methods to improve biocompatibility of nanobeads involving covalent coupling of hydrophilic surfaces

Designed and conducted experiments in the development of fluorescent nanobead -antibody protein conjugation chemistry for cytokine identification and localization (Intracellular and Cell Surface Staining projects).Evaluated several combinations of modified styrene surfaces and modified target functional groups for direct and indirect coupling after oxidative or reductive chemistry. Conventional and bead ELISA development.Testing of proprietary software developed for digital imaging and quantification of multihued fluorescent nano-particlesEvaluated methods to improve biocompatibility of nanobeads involving covalent coupling of hydrophilic surfaces

Performed experiments in the development of protein biochips.Contributed significant improvements in assay development and process development for cytokine micro array biochipImproved sensitivity of cytokine assay fourfold (4X)Developed novel methods to stabilize tethered proteases on microchip surfaces (for screening Peptides

Performed experiments in the development of protein biochips.Contributed significant improvements in assay development and process development for cytokine micro array biochipImproved sensitivity of cytokine assay fourfold (4X)Developed novel methods to stabilize tethered proteases on microchip surfaces (for screening Peptides

Performed experiments in the development of protein biochips. Contributed significant improvements in assay development and process development for cytokine micro array biochip Improved sensitivity of cytokine assay fourfold (4X) Developed novel methods to stabilize tethered proteases on microchip surfaces (for screening Peptides)

Evaluated pharmaceuticals for dissolution, particle size and purity using HPLC and particle sizer in accordance with GMP and

Evaluated pharmaceuticals for dissolution, particle size and purity using HPLC and particle sizer in accordance with GMP and

Scientist III (proteins, metabolites) 8 years experience Published first native protein purification and kinetic characterization of Adenylosuccinate synthetase and Inhibition by Phosphohydantocidin in plants Characterized mechanism of Inhibition of Enzymes by small molecules from libraries and synthesis by in-house chemists Purified to homogeneity and kinetically characterized various Proteins from crude native source materials and recombinant sources Developed and worked with numerous spectrophotometric, fluorimetric, colorimetric and HPLC enzyme assays Designed multiple enzyme pathway assays, including substrate synthesis, both chemical and enzymatic Implemented, developed, and defended (in scientific project meetings) a biochemical target site involving several enzymes in a biosynthetic pathway. From literature search to design of required enzyme assays and substrate synthesis (in collaboration with the chemistry department). Sought out collaborators and justified creation of the new project involving multiple departments. Developed numerous HPLC methods for enzyme assay and metabolite assay (RP, HIC, Ion exchange/pairing).

Developed and implemented protein purification schemes for many native proteins from natural sources. I then developed assays for the screening of synthesized (small molecules) and natural compounds with the purified enzymes. I also used radiolabels to trace compounds through metabolism

Developed new ELISA applications for robotic laboratory workstations (Biomek and GenELISA)Provided Tech support for robotic workstation-software/hardware, both telephone and in customer laboratoriesWrote technical publications for robotic workstationPartnered with the Marketing, Sales, Engineering, and Software departments to develop new uses for the laboratory roboticsDelivered presentations to university and commercial laboratories on the use of the robotics systemsWorked with Engineering and Software departments to troubleshoot new software and hardware, and to develop new attachments, accessories for the robotic workstation

Developed new ELISA applications for robotic laboratory workstations (Biomek and GenELISA)Provided Tech support for robotic workstation-software/hardware, both telephone and in customer laboratoriesWrote technical publications for robotic workstationPartnered with the Marketing, Sales, Engineering, and Software departments to develop new uses for the laboratory roboticsDelivered presentations to university and commercial laboratories on the use of the robotics systemsWorked with Engineering and Software departments to troubleshoot (alpha and beta testing) new software and hardware, and to develop new attachments, accessories for the robotic workstation

Research Assoc I, II, III (Process Development)Assisted in development of protein purification schemes, analyzed fractions with ELISATrained/ assisted production purification team in scale up in pilot plantPerformed small batch protein (1L) production experiments and scale up to 100LPerformed QC for recombinant protein pilot plant lots

Research Assoc I, II, III (Process Development)Assisted in development of protein purification schemes, analyzed fractions with ELISATrained/ assisted production purification team in scale up in pilot plantPerformed small batch protein (1L) production experiments and scale up to 100LPerformed QC for recombinant protein pilot plant lots