Protein Biochemist/Molecular Biologist

Neil A. Johnson

Email: *******@*****.***

Technical Skills and Familiar Instrumentation_______________________

Molecular Biology Techniques:

Polymerase Chain Reaction (PCR), site-directed mutagenesis, overlap-extension PCR, Real-time PCR, colony PCR. Molecular cloning and quantification of DNA, DNA digestion, DNA ligation, agarose gel electrophoresis, DNA sequencing, Isolation of plasmid DNA from yeast and bacteria. Transformation of E. coli and yeast, electroporation, transient transfection of human cell siRNA design and RNA interference (RNAi) techniques.

Cell Culturing and Protein Expression Techniques:

Cell culturing and large to small scale transient expression of recombinant proteins in E. coli bacteria, Pichia pastoris and Saccharomyces cerevisiae yeast and mammalian HEK293 cells (microgram to gram scale). Experience with Drosophila S2, Spodoptera frugiperda Sf9 and Hi5 insect cell expression systems.

Protein Purification Techniques:

Protein and peptide purification using affinity, size exclusion, ion exchange, hydrophobic interaction and reverse phase chromatography (FPLC, HPLC and gravity). Differential centrifugation and equilibrium sedimentation. Processing and purification of endogenous protein from whole tissues. Protein formulation and stabilization. Establishment and maintenance of an endotoxin-free purification environment, endotoxin determination (LAL assay, EndoSafe PTS Meter) and endotoxin removal. Dialysis, de-salting and buffer exchange.

Protein Characterization and Protein Biochemistry Techniques:

SDS-PAGE, Western Blot and NativePAGE. Protein quantification by Bradford assay, BCA assay, Lowery Assay and densitometry. Determination of protein oligomeric state and post-translational modification using non-destructive technologies - dynamic light scattering (DLS), Analytical Size Exclusion Chromatography (AnSEC) and Multi-Angle Light Scattering (MALS). Mass spectrometry. Enzymatic modification and chemical cross-linking of proteins and peptides. Kinetic and ligand binding assay design (i.e. Biacore-based kinetic assays, Solution equilibrium titration, ATP-hydrolysis, radioactive Ca45-flux, fluorescent nucleotide binding and intrinsic absorption heme determination assays). Kinetic, protein quantification and antibody epitope binning assays using the ForteBio Octet Red. Protein : protein interaction and protein pull-down assays.

Vapor diffusion protein crystallization methods.

Familiar instrumentation:

PTI 814 fluoremeter, hemacytometer, microscopy, Nanodrop ND-1000, PTC-200 Peltier DNA thermocycler, DNA sequencer, DNA and protein electrophoresis apparatus, Western blotting apparatus, Bio-Rad electroporator, various centrifuges and

ultracentrifuges, French Press, high pressure homogenizer (HPH), bead mill homogenizer, UV/VIS SpecraMax Plus Spectrophotomer and 96-well plate reader, AKTA FPLC, Waters HPLC, Wyatt Multi-Angle Light Scattering (MALS), Kodak and

Fuji gel imagers, LI-COR Odyssey Imager, Biacore T100 and T200, ForteBio Octet Red, Vi-Cell Analyzer, Q-TOF Ultima Mass Spectrometer, Iatroscan Mk-6s TLC-FID, Wyatt DynoPro Plate Reader (DLS), Charles River Labs EndoSafe PTS Meter, scintillation counter.

Computing and Automation:

Microsoft Word, Excel, Power Point, KaleidaGraph and Adobe Photoshop. Deep-viewer and Pymol protein modeling programs.SoftMax Pro, Unicorn 5.01, Empower, Dynamics v6, MassLynx v4 and Image j

Work Experience___________________________________________________

Novartis Institutes for BioMedical Research: 2009 – Present

Scientific Associate II

Work within a biologics group focused on discovering, designing and generating novel biologic molecules for the treatment of cardiovascular and metabolic diseases. Design, and perform the molecular biology to build, bacterial, yeast, insect and human cloning and expression plasmids. Culture bacterial, yeast, insect and human cell lines. Furthermore, I develop and optimize culturing and expression conditions to produce target and therapeutic proteins in these systems at the microgram to gram scale. Develop and optimize strategies to purify membrane, soluble and secreted proteins using various purification techniques. Additionally, I perform quality control analyses on all purified protein products. Characterize peptides and proteins using various characterization technologies and functional assays. Keep detailed lab notebooks, analyze data, write reports, read patents/primary literature and present findings to colleagues and superiors. Seek new and impactful technologies and instrumentation for my group. I perform validation experiments on adopted technologies and instrumentation, facilitating integration into my group’s standard work-flow.

United States Army (active duty): 1997 - 2001

Airborne Infantryman (Paratrooper), Radio Telephone Operator (RTO)

I managed ~$750,000 worth of sensitive communications equipment.

Education__________________________________________________________

Graduate

Illinois State University, Normal, Illinois: fall, 2006 – spring, 2009

M.S. in Biology / Biotechnology

Cumulative graduate GPA = 3.88 / 4.00

Undergraduate

Illinois State University, Normal, Illinois: fall, 2001 – spring, 2006

B.S. in Biochemistry and Molecular Biology

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