John T. Garbutt

510-***-****

adkyuz@r.postjobfree.com

OBJECTIVE

To do strain/cell line research and Process Development leading to development of biological diagnostics and therapeutics

SUMMARY

I am a Laboratory Scientist with experience growing both microorganisms and mammalian cells to express a variety of recombinant proteins. My strengths are the ability to study and determine what chemistry, conditions and parameters that cells need to achieve optimum growth, expression and function

PROCESS DEVELOPMENT CELL CULTURE SKILLS

Mammalian Cell Culture – Growth of several CHO, HEK293, & Hybridoma cell lines to express a variety of recombinant proteins. Antibodies, growth factors and viruses.

Transformation with expression vectors and growth of E.coli to express a variety of recombinant proteins

Designed and implemented experiments to compare and screen media that supported optimal growth and expression of antibodies and growth factors DOE-JMP

HPLC using affinity Column chromatography to purify Connective Tissue Growth Factor (CTGF) and antibodies

a)Sephadex Affinity column for purification of CTGF

b)Mab Select SURE for purification of antibodies

Recombinant protein purification/fractionation by affinity, ion exchange, and size exclusion chromatography, UV-Vis analysis

SUPPORT SKILLS & EXPERIENCE

Basic molecular biology skills such as DNA & RNA extraction, plasmid production and purification, DNA Ligation, PCR amplification, qPCR

Cell Line Development - Transfection of mammalian cell lines with DNA vectors to express antibodies.

Use of HPLC systems (Protein A and SEC) to purify and measure antibodies

Use of Octet and Biacore to quantitate and measure protein binding

Protein expression detection techniques - SDS-PAGE, Western Blot, Elisa

EDUCATION

Master of Science – Microbiology/Molecular Biology

University of Wyoming, Laramie WY

PROFESSIONAL EXPERIENCE

Fibrogen, San Francisco, CA Contract January2020-Present

Senior Research Associate, Cell Line Development and Protein Expression

Protein concentration and buffer exchange by tangential-flow filtration

HPLC using affinity Column chromatography to purify Connective Tissue Growth Factor (CTGF) and antibodies

c)Sephadex Affinity column for purification of CTGF

d)Mab Select SURE for purification of antibodies

Recombinant protein purification/fractionation by affinity, ion exchange, and size exclusion chromatography

SDS-PAGE and Western blotting of samples

ELISA analysis of conditioned media from cells to titer antibody levels as well as other marker proteins.

Fluidigm, South San Francisco, CA Contract February2019-December2019

Senior Reagent Manufacturing Associate, Proteomics Reagents Operations

Manufacturing of Isotopic-metal conjugated antibodies, QC testing of raw materials, in-process materials and finished goods for release. Subject Matter Expert in the development and testing of new procedures

Performed biological experiments (cell culturing, cell isolation, cell staining)

Maintenance and sustaining of the cell bank

Performed product testing using standard ICP-MS and novel Mass Cytometry technology on CyTOF instruments

Smartphage, South San Francisco, CA Consultant Jun2018- December 2018

Production of bacteriophage against Cutibacterium acnes (clinical bacterium causing acne)

Guidance and assistance to make Master Cell Bank and Working Cell Bank

Experiments to define optimum concentrations of components in growth medium to achieve optimal growth of bacterial culture (Increased growth 4X)

Testing MOI (Multiplicity of Infection) for amount of bacteriophage to infect host bacterium with to produce maximum yield of bacteriophage (Increased two logs)

Provided a bench scale clarification and filtration system that increased yield of bacteriophage 3-4X (Centrifuge, PP3 1.2um-0.65um, XLG 0.2um)

Selection of TFF system for filtration, concentration and buffer exchange (GE AKTA FLUX)

Write-up of protocol for CMO to use to scale process from ml at bench to Liters in bioreactor

BD Biosciences, San Jose, CA Contract Jan2018-Jun2018

System Verification and Validation Scientist

Execution of verification activities throughout all phases of the FACS product development process

Assist in design of test methods and test strategy

Development and execution of formal protocols

Analysis and interpretation of results

Communication of results and their impact through written reports, and interactive discussions

Conducted experiments in flow cytometry to define best laboratory procedures and study designs

JGB Bio Pharma, Mountain View, CA Short Term Contract July-Aug 2017

Quality Control Analyst

Growth of Modified Bone Marrow-Derived Mesenchymal Stem Cells for analysis

Receipt and catalog of frozen clinical samples of stem cells

Thaw of stem cells and expansion for tesing

Staining of stem cells for FACS analysis and PCR

Ignite Immunotherapy, Alameda, CA Short Term Contract March 2017

Research Associate II, Start-Up Lab Operations

Assist in the procurement, set-up of equipment, stock supplies and cell banking

Collect quotes on equipment and arrange for delivery

Oversee and install laboratory equipment

Receive and catalog shipments of cell & virus cultures

Thaw expand cell culture and research cell banking (RCB)

Infect cell for virus propagation and RCB

Merck Research laboratories, Palo Alto, CA Contract March2016-Jan2017

Technical Specialist, Antibody Generation

Growth of Hybridoma cell lines to produce antibodies for characterization and scale up

Expansion of screened hybridoma cell lines to produce antibodies for characterization, analytical testing, and cell banking

Growth of rat and mouse hybridoma cell lines in T-Flask, Spinner tube, shake flask, Integra bioreactor and small scale bioreactor to produce antibodies for epitope binning, sequencing and kinetic analysis

Designed experiments for production, work flow, quantization by octet384 and initial purification. Documentation of work & results in database and electronic notebook with presentation of analyzed data to various groups

Portola Pharmaceuticals, South San Francisco, CA Contract April2015-Jan2016

Associate Scientist, Biology

Cell line development in CHO to express Andexanet by transfection and electroporation

Optimized NEON electroporation conditions for vectors

Successfully generated CHO-S culture that had stable integration of linearized plasmid DNA

Selection of 10X producing cell line pools for Andexanet

Designed and planned out bioreactor laboratory

Selection of bioreactor system and supporting equipment

Configuration, Set-Up, Operation and harvest of 10L stirred tank

Documentation of data into reports, Power Point Presentations

University of California-San Francisco-Mission Bay July2013- April2015

Staff Research Associate III, BioNanotechnology

Production of M13 bacteriophage from Lab-Scale bioreactors to supply ssDNA to fold and construct multiple DNA nanostructures.

Developed a bench-scale process to produce M13 from 2L bioreactors

Developed a recovery and purification process for M13 bacteriophage

Extract and purify M13 DNA

Assist in the folding of M13 template DNA into “DNA Origami Nanostructures”

Assisted in molecular biology, protein expression and strain development

Developed a qPCR assay for quantification of M13 DNA

Medimmune LLC, Santa Clara, CA July2010-Dec2012

Associate Scientist II, Vaccine Clinical Production

Process development and cGMP clinical manufacturing of Live Attenuated Influenza Vaccine (LAIV). Establish and prepare Working Cell Bank, Virus Working Banks.

Thaw of WCB adherent MDCK & Vero cells

Expansion of cells in T-flasks and roller bottles with determination of parameters and timing of culture to ensure optimum growth

Set-Up, inoculation, control with data logging of 50L Single Use Bioreactors SUB and harvest of virus

Preparation of Master Production Records (MPR), Technical Reports and presentations

Henlius Biopharmaceuticals, Inc. Fremont, CA Oct2009-June 2010

Consultant, Process Sciences

Process development of cell culture systems expressing biosimilars and biobetters (mabs and recombinant proteins) for malignant tumors and auto-immune diseases.

Recommended make, model and scale of bioreactor system for production of biosimilars from CHO cell culture

Arranged for facilities to support bioreactor operations (air compressor, gas cylinders and manifold controller)

Coordinated Installation of bioreactor systems

Training of personnel to set-up, operate, sample and harvest bioreactor cultures

Recommended and acquired off-line equipment for analysis of metabolites and counting of cells ( NOVA FLEX and Vicell)

Sutro BioPharma Inc, South San Francisco, CA Contract Dec 2008-July 2009

Senior Research Associate, Process Sciences

Process development of pilot plant scale fed-batch fermentation of E.coli and conversion into cytoplasmic extract for cell free protein production. Research on cell-free extract protein production technology.

Operation of bioreactors at 1L-10L-200L for growth of E.coli

Homogenization of E.coli cells to form extract

Disc-Stack Centrifugation to differentially separate extract into active component

Running of assays for measuring growth and production of recombinant proteins

Neugenesis Corporation, Burlingame, CA Contract Sept 2007 – April 2008

Fermentation Scientist

Process development for bioreactor growth of filamentous fungus Neurospra Crassa for production of recombinant proteins and antibodies

Set up, operation and harvest of Sartorius stirred tank (3L) bioreactors

Processing of bioreactor time course samples for wcw and dcw measurement of accumulated cell mass

Off-line analysis of glucose consumed and metabolites generated for media development

Feeding of nutrients for fed-batch bioreactor process to drive expression of proteins

Analysis of expressed proteins by SDS-PAGE and western blot

Preparation of technical reports and presentation of results to staff

Berlex Biosciences, Richmond, CA March2005 - April2007

Associate Scientist III, Upstream Process Development

Cell Culture Process Development - Growth of mammalian cells to produce adenovirus, growth factors and antibodies.

Design and implementation of experiments to compare and screen cell culture media that would support optimal growth and expression of recombinant proteins from several different cell lines.

Batch Bioreactor Process Development for the production of recombinant proteins. Determination of Process effects of pH ranges and shifts, dissolved oxygen setpoints, inoculation, seed train and recovery.

Developed Fed-Batch Bioreactor Process that improved yield of antibody 5X over batch process by tailoring a feed strategy that optimized use of substrate with minimum accumulation of byproducts

Scale-up of Fed-Batch process from 3L to 1000L and use of scale-down methodology to trouble shoot the fed-batch process to fit schedule and capacity of GMP facility

Berlex Biosciences, Richmond, CA April2002 – March2005

Associate Scientist II, Upstream Process Development

Cell Culture Process Development - Growth of mammalian cells to produce adenovirus, growth factors and antibodies.

Served on multiple project teams that Coordinated with Research and Development to deliver timely produced materials in support of multiple projects

Wrote numerous technical reports for process development, process transfer and SOP establishment for GMP manufacturing

Data from studies was analyzed with results presented in departmental meetings and scientific conferences

Use of HPLC systems (ProteinA and Size Exclusion Chromatography) to purify and measure antibodies

Berlex Biosciences, Richmond, CA February1999-April2002

Associate Scientist I, Upstream Process Development

Cell Culture Process Development - Growth of mammalian cells to produce adenovirus for Gene Therapy for Myocardial Ischemia

Shake Flask studies to determine optimal Multiplicity of Infection (MOI)

Shake Flask studies to select optimal media for growth of host cell and infection medium to produce virus

Preparation of cell banks (RCB) of HEK293 and Perc.6

Perfusion Bioreactor process development to produce high viability, high density inoculums to seed adenovirus production bioreactors

Continuous Batch bioreactor process development for production of recombinant adenovirus. Selection of optimum pH, temperature and dissolved oxygen. MOI

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