Jeffrey Johnson

Summary

Eighteen years of industry and academic experience in biological and small molecule drug development.Expertise with a diversity of biological research applications including eukaryotic/prokaryotic protein expression, antigen and antibody preclinical validation, small molecule function and formulations, surface plasmon resonance, signal transduction and in vitro assay development, etc.. Comprehensive experience with both transient and stable mammalian cell culture protein expression, vector design, target validation, and molecular biology. Innovative, adaptable, resourceful and effective at advancing project milestones.Excellent problem solver.Works proficiently in both independent and team oriented projects. Excels in dynamic laboratory environments.

Objective

Challenging market conditions increasingly require biotechnology companies to expeditiously evaluate therapeutic candidates. I have the experience and expertise to screen, validate, and manufacture preclinical biologics across a spectrum of hosts and systems.  I will pinpoint and execute those experiments required to identify product leads and navigate them efficiently through the R & D process.

Work History

Work History

Research Assistant, Robert Hinrichsen Laboratory

Fred Hutchinson Cancer Research Center

¨Assisted with projects related to the discovery and cloning of a novel PP2A gene from Paramecium.

2007 - Jan 2012

Scientist, Cell Line Development and Molecular Biology

Allozyne, Inc.

¨  Developed a mammalian tissue culture expression platform in HEK293 c18 and CHO cells (DG44 and CHO-S) for site-specific incorporation of nonnatural amino acids (NNAA) into therapeutic IgGs via amber suppression by an orthogonal tRNA-aaRS pair for downstream toxin conjugation (ADC’s) by click chemistry. Demonstrated high yield IgG-NNAA production with manufacturing scale feasibility.

¨Developed an orthogonal tRNA-aaRS pair for site-specific incorporation of NNAA’s in E. coli for the manufacturing of recombinant therapeutic proteins.

¨Expressed, purified, and characterized antigens, antibodies, and antibody fragments used for downstream product development.

¨Developed surface plasmon resonance assays for product candidate validation/affinity determinations.

¨Developed ELISAs used for quantitation and pharmacokinetic analysis of therapeutic candidates.

2005 - 2007

Scientist I

Ikaria, Inc.

¨Developed flow cytometry assays to assess the efficacy of Ikaria’s technology on blood platelet storage.

¨Expanded Ikaria’s compound portfolio via an in vivo lethal hypoxia SAR screen.

¨Developed a multiwell plate oxygen consumption assay for characterizing compound potency.

¨Characterized a panel of excipients for lead compound stability and formulations development.

¨Optimized manufacturing process of lead compound for In vivo toxicology studies.

¨Evaluated stability of lead compound for FDA required In vitro toxicology studies.

¨Developed a rapid tissue cryopreservation method in rats for measuring lead compound metabolites.

Mar 2005 - May 2005

Senior Research Associate, mAb Development

Seattle Genetics

¨Developed a functional assay for screening a panel of mAbs coupled with a toxin conjugate.

¨Established a flow cytometry quantitative antigen density  assay for a multiple myeloma associated protein.

1997 - 2004

Senior Research Associate, mAb Development

Corixa Corp.

¨Expressed and validated Corixa’s proprietary tumor antigens in mammalian

       culture models for vaccine, therapeutic and diagnostic antibody development.

¨Evaluated antibody binding and function against tumor antigens for therapeutic

       and diagnostic development.

¨Produced hybridoma’s, recombinant antibodies, and cancer vaccine proteins in

       mammalian expression systems, including CHO and human cell lines.

¨Cloned, expressed, and characterized a modified Her2/neu CHO expressed

      protein vaccine partnered with GSK.

¨Expressed and evaluated multiple forms of an atypical apoptosis inducing anti- 

      GM2 monoclonal antibody and demonstrated that cytotoxicty was dependent on

       specific domains within the Fc.

¨Established HEK and CHO expression models used for antigen and antibody

       development.

¨Grew and maintained 30+ adherent and suspension cell lines from multiple 

      species and tissue types.

¨Developed an antibody dependent cell-mediated cytotoxicity (ADCC) assay using

       Lactate Dehydrogenase (LDH) as the cytotoxicity indicator.

¨Expression Cloned and characterized a novel anti-CD9 apoptosis inducing

       antibody.

¨Trained Research Assistant and supervised project progress for 6 months

    (temporary assignment).

¨Produced large quantities of mIL-4 using an inducible insect cell culture system 

       resulting in significant cost savings to the company.

1994 - 1997

Research Technician II, Jonathon A. Cooper Laboratory

Fred Hutchinson Cancer Research Center

¨Characterized and imaged (using 3-D Deltavision microscopy) a novel kinase 

      (Mnk 1) involved in the regulation of translation.

¨Established a reproducible focus formation assay and analyzed the

       transformational activity of mutations of Ha-Ras.

¨Managed a 10 person laboratory.

Education

Education

University of WA

Howard Hughes Summer Research Internship, John Edwards Lab, Zoology,  1992

Sep 1989 - May 1993

BSc, Magna Cum Laude

Seattle Pacific University

Magna Cum Laude

Skills

Skills

Other:

Ø  Toxic gas handling/glove box use Ø  In vivo experience including mouse/rat drug delivery, tissue         harvesting,  glucose monitoring, etc.

Molecular Biology

Ø  Recombinant DNA/subcloning/RNA isolation/expression Ø  PCR/Rev. Transcript.-PCR Ø  Site-specific mutagenesis

Cell Biology/Biochemistry/Analytical

Ø Multicolor flow cytometry, quantitative flow cytometry Ø Surface Plasmon Resonance/affinity kinetics Ø Antibody internalization assays Ø Cytotoxicity  and metabolism assays (Resazurin, MTS, Cell Titer Glo) Ø ELISA Ø SDS-PAGE/ densitometry/Immunoblotting/ Protein detection assays (ECL, etc.) Ø Protein labeling (Alexa, biotin, etc.) Ø Cellular fractionation and subcellular protein localization assays Ø Apoptosis/proliferation/metabolic assays (BrdU, Annexin, CaspaTag, anti-single stranded DNA antibody, PI, MTS, LDH, oxygen consumption) Ø Immunofluorescence microscopy (including Confocal and Deltavision 3-D microscopy) Ø Affinity protein purification (IMAC,Protein A/G, immunoprecipitation, etc.) Ø Fluoroblock Matrigel inserts Ø Fluorometry/luminometry Ø Multiple Epitope tags and reporter systems (His, Biotin Ligase, DSRed II, GFP, eGFP, hrGFP, FLAG, GST, HA, Src, Myc, Secreted AP, LDH, b-Gal)

Cell Culture

Ø Multiple protein expression systems (mammalian, insect, yeast, and E. coli) Ø Transient transfection optimization and stable cell line development (+/- serum, shake flasks, semi-solid and limited dilution cloning, etc.) Ø Pol. III driven stable tRNA expression Ø Integra bioreactor/ Cell factory/ Unisyn Cell Pharm hollow fiber bioreactor Ø Focus formation transformation assays Ø ADCC Ø Microcarriers Ø Colloidal gold migration assays Ø Hybridoma development