Quality Control Customer Service

SKILLS SUMMARY

Professional Cell Culture and Fermentation Specialist who develops and

optimizes Bioprocesses for the Production of Recombinant Proteins: Protein

Kinases, Cytokines, Immunogens, and Industrial Chemicals: 1,4-Butanediol

(BDO).

My experience is with various host cells including, Insect, Mammalian, E-

coli, CHO and Yeast cells (Pichia).

I am flexible team player and leader who takes initiative, trouble shoots,

and identifies solutions to resolve problems for process improvements and

effectiveness. I truly enjoy the science of processing living cells to

produce and optimize products that will help humanity. Skills include:

Baculovirus Expression Vector Quality Control

System

Insect Cell Culture Elisa Assays

Mammalian Cell Culture Immunohistochemistry Staining

Bioreactors, Fermentors Excellent Aseptic Technique

Media Screening & Optimizations FACS

Virus Production Multi-Tasking, Dependable Team

Player

Process Development and Engineering Excellent Problem

Solver/Troubleshooter

PCR, Westerns & SDS Page Gels Customer Service

PROFESSIONAL EXPERIENCE

eBioscience- San Diego, CA - Molecular Biology Dept.

Research Associate II (Sept. 2008- Present)

. Production of Recombinant Proteins for Researchers using the Baculovirus

Expression Vector System (BEVS).

. Perform Co-transfection of transfer vector with linearized baculovirus

DNA into insect cells to generate Recombinant Virus stocks.

. Perform End-Point Dilutions for the isolation of Single Virus Colonies

that express GFP. The clones are then scaled-up and amplified to produce

High Titer Virus Stock that > 99% Recombinant.

. Amplify Recombinant Virus stocks in SF9 host cells.

. Perform Small scale optimizations in shake flasks to test different

media's, cell density, virus volumes (M.O.I's) at infection, cell types,

Harvest times etc. to maximize protein production.

. Perform Concentration of conditioned media, prepare Buffers and purify

Immunogens.

. Consistently Perform 5, 10 & 20L Protein Expressions in insect cells

using Rollerbottles, and disposable Wave Bioreactors using high titer

Recombinant Virus stocks.

. Maintain SF9 insect cells for Virus Amplification and Protein Production.

. Perform the following Molecular Biology techniques: DNA extraction and

Purification using Qiaquick PCR Purification Kit; Amplification of the

Baculovirus DNA using PCR; Agarose Gel Electrophoresis with Ethidium

Bromide to visualize and separate the Molecules by size, Western blots

using One-Step Western Complete Kit with a Mouse Primary Antibody by

Genscript to detect and visually quantify the expression of our Proteins.

. Perform work independently and under minimal guidance and supervision.

. Co-management of the molecular biology lab.

Nexbio Inc.- San Diego, CA - Process Development Group.

(3 month assignment as an Individual Contractor)

Senior Research Associate (2007- 2008)

. Responsible for the Fed-Batch production of Proteins (Enzymes) using E-

coli cells in 10L & 20L Fermentors (Steam in Place) from New Brunswick

Scientific. Documented using Batch records.

. Media preparation and filtration. Also, analysis of key nutrients and

metabolites in cell culture and fermentation growth media using a Nova

BioProfile Chemistry Analyzer.

Genomatica Inc.- San Diego, CA - Microbiology/Fermentation Group.

(Contracted through Kelly Scientific Resources, 13 month assignment)

Fermentation Specialist (2006-2007)

. Conducted laboratory experiments to support Genomatica's strain

engineering programs.

. Focused was on optimizing microbial strains, media, High-Throughput

Fermentations and adaptive evolution of Microbes for the production of

renewable chemicals (i.e 1,4-Butanediol).

. Experiments were conducted in shake flasks which were then scaled up into

multiple Batch or Chemostat Fermentation runs (Bio-Factories) using the

Bioflow 3000 (5L bioreactor), the Bioflow 110 (1.3L bioreactor) from New

Brunswick Scientific and Biostat Bplus twin benchtop

bioreactors/fermentors from Sartorius Stedim Biotech (2x 3L bioreactors)

with the ultimate goal of improving productivities, product yields and

overall process economics.

. Activities included but not limited to: growth of microbial cultures in

shake-flasks, batch fermentations, and chemostat fermentations;

Microbiological techniques: media preparation, plate streaking and

counting, cryopreservation, aseptic techniques, and serial dilutions.

. Expertise in Anaerobic and Aerobic cell culture and fermentation

involving E-coli bacteria and Yeast cells for the production of

metabolites and recombinant proteins.

. Analysis of metabolites (Glucose and Ethanol) using the Analox "Micro-

Stat" GL6 Analyzer.

. Able to work effectively with minimal supervision and as a team member

with the entire company.

. Excellent project management by multi-tasking, time management, weekly

spreadsheets/graphs, consultation with piers, organization, problem

solving, technical knowledge and experience.

PacificGMP & Arizeke Pharmaceuticals - San Diego, CA - Process Development

Group.

(Contracted through Payrolling.com)

Senior Production Associate (2004 - 2005)

. Produced antibodies using HK-PEG-1 hybridoma cell lines and fusion

proteins using mammalian A549 and HT1080 cell lines in Cell Factories,

Flasks, Spinners, Cell Cube and Wave Bioreactors.

. Performed transient transfections, and adapted adherent cells to

suspension cells and serum free.

. Selection and maintenance of eukaryotic cells transfected with the

hygromycin B resistance gene.

Pfizer Global R&D - San Diego, CA - Molecular Biology Dept. - Baculovirus

Fermentation Group.

Senior Associate Scientist (2003 -2004)

Associate Scientist (1998- 2003)

. Small (<5-Liters) and large-scale recombinant protein (Enzymes)

production using Insect Cells and the Baculovirus Expression Vector

System in "Wave Bioreactors" and 18-Liter Applikon Bioreactors.

. Monitored critical parameters in the Bioreactors such as: temperature,

cell density, DO, pH, and glucose uptake.

. Large-scale amplifications of recombinant viral stocks in Roller

Bottles/Shake Flasks.

. Transfection and single plaque isolation used for virus amplification.

. Process optimization studies involving insect cell host (SF9 & Hi5), cell

density at infection, multiplicity at infection (MOI), and post-infection

harvest time to maximize yield of recombinant protein.

. Quality control of host cells, media, serum and recombinant viral stocks.

. Cell thawing and freezing of master and working cell banks (inoculums)

used for seed train process.

. Cleaning, preparation, maintenance and sterilization of the production

fermentor and accessories.

. Prioritized, organized, and followed-up on the protein needs for all

internal customer groups.

. Negotiated contracts involving equipment, media and supplies that saved

the company thousands of dollars, and produced a consistent "high

quality" lot of raw materials.

. Initiated and helped designed fermentation databases; a tool for

scientists to order services, track/retrieve information on their

projects in real time, saving the company thousands of dollars.

. Wrote SOPs, reports, data entry in notebooks and databases, and presented

scientific data to group.

. Executed SDS-PAGE gels to assess protein expression and purity as in-

process controls.

. Supervised lab personnel by providing training, guidance, and

encouragement for professional development.

Immusol Inc. - San Diego, CA - Stem Cell and Process Development Dept.

Cell Culture Technologist (1996-1997)

. Processed umbilical cord blood for the isolation of mononuclear cells.

. Isolated CD34+ hematopoietic stem cells using a miniMax column

bioseparation kit.

. Maintained numerous cell lines in T-Flasks, including CHO, A549, HEK 293

and HT1080.

. Created in-house guidelines for culturing adherent/suspension cells.

. Established banking system to track cryovials, thus saving time and

money.

Dako Corporation - Carpentaria, CA - Biochemical Production Dept.

Cell Culture Technologist (1995-1996)

. Established and banked in-house supply of master seed and working seed

stocks.

. Assured quality of master and working seed stocks released to cell

production by conducting quality control testing with

immunohistochemistry staining and Elisa assays.

. Acted as liaison between research and development (R&D) and BCP in the

transfer of new monoclonal antiserum from hybridoma cell lines.

. Maintained GMP documentation records per company standards and government

regulations.

. Prepared large volume dilutions of immunochemicals and enzyme reagents

for kit assembly.

. Assured quality of formulations released to kit assembly by conducting in-

process quality control (IPQC).

. Screened untested tissue blocks for the BCP QC tissue bank.

. Maintained BCP raw materials inventory records per kit lot order.

Systemix Inc. - Palo Alto, CA - Cell Expansion Development Dept.

Research Associate I/II - Cell Products Development Associate (1994)

. Maintained long-term in vitro progenitor cell cultures on a small-scale

through clinical-scale basis.

. Performed techniques for cell analysis including cell harvesting,

fluorescent staining of hematopoietic stem cells, FACS, and methyl

cellulose assay to support bioreactor and plate experiments.

. Developed and performed experiments in bioreactors, plates and flasks to

determine the optimum cell inoculum density, cytokine

variation/concentration, and length of stem cell expansion.

La Jolla Institute for Allergy and Immunology - La Jolla, CA

Research Technician I (1992-1993)

. Maintained a long-term cell culture of mononuclear cells of umbilical

cord blood in the presence of Recombinant human C-kit ligand (stem cell

factor) to develop functionally active human mast cells.

. Maintained a long-term cell culture of bone marrow derived mouse mast

cells (BMMC) in the presence of IL-3 to develop functionally active mouse

mast cells of high purity (>90%).

EDUCATION

. Bachelor of Science in Applied Arts and Sciences Microbiology, San

Diego State University

. Associate of Arts, Mesa College, San Diego, CA

. Business and regulatory practices class (cGMP's) from Mira Costa

College, 2004

. Bioprocess and cell culture class from Mira Costa College, 2005

PUBLICATIONS

. Chen Ping, Luo Chun, Deng Yali, Ryan Kevin, Register James, Margosiak

Stephen, Tempczyk-Russell Anna, Nguyen Binh, Myers Pamela, Lundgren

Karen, Kan Chen-Chen, O'Connor Patrick M. 1.7A Crystal Structure of human

Cell Cycle Checkpoint Kinase CHK1: Implication of CHK1 regulation, Cell,

Volume 100, Issue 6, 17 March 2000, Pages 681-692.

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