Engineering Supervisor

FANG FANG, Ph.D.

Phone: 404-***-****

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

Address: ***** ***** *** *, *********, WA, 98133

USA Status: Permanent resident

SUMMARY OF QUALIFICATIONS

Solid theoretical background and hands-on experiences in biochemistry, molecular biology and genetics in various biological systems including E. coli, yeast, and mammalian cells.

Intensive experience in the allergy test field including antigen extract preparation, antigen QC, immunoassay development, validation and QC.

Excellent skills on bioassays such as ELISA, Luminex, beta-gal and dual luciferase assay.

Expertise on protein engineering such as gene assembly, library construction and high throughput screening.

Extraordinary DNA construction capability. Commendations as “the best DNA cloner in the past thirty years in the lab” by Dr. Suzanne Sandmeyer.

Proficient in yeast genetic manipulation and strain metabolic engineering.

Team oriented for solving complex biological challenges and excellent communication.

Excellent collaborations with cross-disciplinary team members (chemists and bioinformatics researchers).

Lab management: economical and efficient utilization of resources; instruction of more than 10 research personnel.

PROFESSIONAL EXPERIENCE

US BioTek, Shoreline, WA

Techinical supervisor in Immunology department, Nov, 2015-present

Job description: Supervises and maintains laboratory facilities for routine testing, experiments and research in new procedures. Coordinates and plans laboratory tests and conducts experiments for approved new projects. Supervises and provides technical guidance to staff and teaching assistants concerning pre- and post-experiment preparations and processes. Ensures compliance with CLIA, COLA and OSHA during laboratory operations.

Select test methodology that is appropriate for the clinical use of the test results.

Verify the test procedures performed and establish laboratory's test performance characteristics, including the precision and accuracy of each test and test system.

Resolve technical problems and ensure that remedial actions are taken whenever test systems deviate from the laboratory's established performance specifications.

Establishes the acceptable levels of analytic performance, and ensures these levels are maintained throughout the testing process for pre-analytic, analytic, and post-analytic phases of testing.

Ensuring that patient test results are not reported until all corrective actions have been taken and the test system is functioning properly.

Establish a quality control program appropriate for the testing performed and establish the parameters for acceptable levels of analytic performance and ensuring that these levels are maintained throughout the entire testing process from the initial receipt of the specimen, through sample analysis and reporting of test results.

Ensure that remedial actions are taken and documented whenever significant deviations from the laboratory’s established performance characteristics are identified, and patient test results are reported only when the system is functioning properly.

US BioTek, Shoreline, WA

Scientist in R&D, Feb, 2014-Oct, 2015

Job description: Optimize and develop allergy test conditions including colorimetric assay (ELISA) and chemiluminescent assay.

Constructed recombinant allergens and tested their activities..

Optimized food allergy tests mainly focusing on ELISA assay to assess IgE, IgG and IgA antibodies. The optimization included sensitivity improvement, conjugate selection and standard curve application.

Carried out the validation of IgA ELISA assay, the conditions such as LOD, LOQ, antibody lot to lot variation and plate stability were checked.

Worked on allergy assay QC.

Improved the quality of food antigens through clean-up methods.

Optimized colorimetric assay (ELISA) for enhancing IgE, IgG and IgA antibody detection sensitivity.

Resolved technical issues for production department.

Department of Biochemistry, University of Washington, WA

Senior Fellow, August, 2012-August, 2013

Supervisor: Dr. Ted Young

Job description: Investigating the interaction of Snf1 and downstream target proteins, with particular emphasis on how the Snf1 kinase pathway influences gene expression.

Statistical analysis of phospho-proteomics data to find out Snf1 target protein candidates.

Construction of Snf1 target genes and expressed them in yeast.

Investigation on Snf1 target protein in yeast via mass spectrometry (MS).

EdeniQ Inc, Visalia, CA

Scientist in R&D, November, 2010-May, 2012

Job description: Yeast metabolic engineering for cellulosic ethanol. Accomplishments for improving sugar conversion into ethanol include:

Constructed strains with over expression of secreted enzymes, while maintaining high activity.

Developed enzymatic assay for testing activity of in-house cellulases and hemicellulases.

Implemented a novel metabolic system into S. cerevisiae by integrating heterologous proteins for sugar utilization and conversion to ethanol.

Adapted strains for high sugar utilization, tolerance for ethanol, inhibitors and acids.

Engineered diploid strains to increase bioprocess efficiency. Tested their effectiveness via growth rate, inhibitor tolerance, protein expression, saccharification and fermentation.

Modified concentration of enzymes based on kinetic experiments, for improved biomass saccharification efficiency. Worked with in-house and commercial cellulases.

Carried out quality assurance of dry yeast for commercialization such as genotyping.

Department of Biological Chemistry, University of California-Irvine, CA Specialist, September, 2008 – November, 2010

Supervisor: Dr. Suzanne Sandmeyer

Job description: (1) Designed and developed novel yeast strains for lignocellulosic biofuels production. (2) Engineered Saccharomyces cerevisiae for short-chain fatty acids production via biorenewable approaches.

Constructed more than 100 plasmids for systematic metabolic engineering in S. cerevisiae; around 300 of pXP plasmids have been purchased by researchers through Addgene.

Manipulated more than 50 genes and knockout in S. cerevisiae.

Constructed xylose reductase (XR) library using DNA self-assembly technique and performed high throughput screening to optimize the solubility of XR.

Engineered S. cerevisiae strains to increase the yield of fatty acids and carried out strain evolution for increasing fatty acid production by library screening.

Identified crucial genes for higher fatty acid production by microarray analysis.

CODA Genomics (now named as Verdezyne), Carlsbad, CA Postdoctoral Researcher in R&D, August, 2007 –August, 2008 Job description: Developed and produced high levels of soluble proteins for external or internal targets of interest from their previously insoluble versions using CODA Genomics’ SolSelect™ Platform Technology.

Implemented hundreds of gene assembly using CODA Genomics’ optimized nucleotide sequences.

Finished over 10 different library constructions and screening for highly soluble proteins.

Conducted soluble mutant identification and protein expression in E. coli.

Modified vectors for optimization of protein expression.

Emory University, Atlanta, GA Graduate Researcher, September, 2001-August, 2007

Supervisor: Dr. David Lynn

Job description: Investigated the mechanism of phenol sensing and transduction of VirA/VirG two-component system in Agrobacterium tumefaciens.

Proved VirA as the direct receptor for phenol sensing, resolving the long-standing controversy on phenol sensor in Agrobaterium.

Constructed various mutants and characterized the mechanism of VirA upon phenol sensing.

Discovered and defined novel functions of VirA linker domain and receiver domain.

Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, China

Graduate Researcher, August, 1998 – June, 2001

Genetically engineered, expressed and characterized novel catalytic antibodies that mimic glutathione peroxidase.

PROFESSIONAL SKILLS

Molecular biology: DNA cloning, PCR (colony PCR, site-directed PCR, Q-PCR and RT-PCR), electrophoresis, southern blot, western blot, etc.

Protein: Protein engineering and expression in E. coli, Agrobaterium and S. cerevisiae. Protein purification with Ni-NTA agarose, DEAE anion-exchange column and S-300 gel filtration

Bioassay: ELISA, Beta-gal, dual luciferase, bacterial two-hybrid assay, enzymatic assay and in vitro/in vivo protein cross-linking.

Yeast genetics: Cultivation, transformation, gene knockout, gene integration, genotype, diploid formation, DNA extraction, RNA extraction, immunoprecipitation (IP), and lipid extraction.

Cell culture: Maintenance of mammalian cell line, siRNA knock down, transfection, transduction, RNA extraction, protein extraction and stable cell line generation.

Analytics: Luminex, MSD, UV-vis, HPLC, GC-MS and Light/fluorescent microscopy.

Computer: Adobe Photoshop, office software, DNA sequence analysis (e.g. Blast, alignment, vector NTI), Protein structure modeling (e.g. Swiss Model), Codon-usage analysis in different organisms and microarray statistical analysis.

EDUCATION

Emory University, Atlanta, GA,

Ph.D. in Molecular Biology, 2007, Advisor: Professor David G. Lynn

Changchun Institute of Applied Chemistry, Chinese Academy of Science, Changchun, China

M.S. in Biochemistry, 2001, Advisor: Professor Daqing Zhao

Beijing Normal University, Beijing, China B.S. in Chemistry, 1998

PUBLICATIONS

Fang, F; Lin, Y; Lynn D.G. A Rhizobium radiobacter histidine kinase can employ both Boolean AND and OR logic gates for initiating pathogenesis. ChemBioChem. (2015), 16: 2183-2190

Fang, F; Stewart T; Zhang J. Biomarkers of Parkinson’s disease. General Methods in Biomarker Research and their Applications. 2015, (1-18).

Braun, K; Vega, S; Dombek, K; Fang, F; et al. Phosphoproteomic analysis identifies proteins involved in transcription-coupled mRNA decay as targets of Snf1 signaling. Sci. Signal. (2014), 7 (333), ra64. [DOI: 10.1126/scisignal.2005000]

Lin, Y; Pierce, D; Fang, F; Lynn D. G. Role of the VirA histidine autokinase of Agrobacterium tumefaciens in the initial steps of pathogenesis. Front Plant Sci. (2014), 5: 195.

Shen, M; Fang, F; Sandmeyer, S; Da Silva, N. Development and Characterization of a Vector Set Allowing Controlled Gene Expression during Systematic Metabolic Engineering of Saccharomyces cerevisiae. Yeast (2012), 29: 495-503.

Fang, F; Salmon, K; Sandmeyer, S et al. A vector set for systematic metabolic engineering in Saccharomyces cerevisiae. Yeast (2011), 28: 123–136. This paper has been evaluated as a top paper in Yeast.

Wise, A.A; Fang, F; Lynn D.G.; Binns, A.N. The receiver domain of hybrid histidine kinase VirA: an enhancing factor for vir gene expression in Agrobacterium tumefaciens. J. Bacteriol (2010), 192: 1534-1542.

Gao, R.; Mukhopadhyay,A.; Fang, F.; Lynn D.G. Constitutive activation of two-component response regulators: Characterization of VirG activation in Agrobacterium tumefaciens. J. Bacteriol. (2006), 188: 5204-5211.

Fang, F, Wang, L., Qi, X., et al, Human catalytic antibodies with glutathione peroxidase activity. Journal of Inorganic Biochemistry. (2001), 85: 301-7.

Qi, X., Wang, L., Fang, F, et al, Studies of soluble human catalytic antibody with glutathione peroxidase activity. Chemical Journal of Chinese Universities. (2001), 22: 1976-78.

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