Research scientist, cell/molecular biology, protein biochemistry
Resume:
CURRICULUM VITAE
Barbara-Anne Martin
** ****** ***, ********, ** 02891
******.*************@*****.***
OVERVIEW:
Scientist with experience in pharmaceutical, academic, and hospital research settings
Technical competency in Immunology, Protein Biochemistry, Cell/Molecular Biology
Able to work independently or as part of a multidisciplinary research team
Strong organizational ability with attention to detail
TECHNICAL SKILLS:
Assay development
Isolation/characterization of dendritic cells to monitor effects of viral infection
Binding assay to characterize dendritic cell antibodies
Coculture assay to detect dendritic cell interaction with T cells
Indirect ELISA for the clotting factor FVIII
Multiplex bead-based assay to detect multiple cytokines in plasma & brain
FACS assay for phenotypic analysis of peripheral blood lymphocytes
In Vitro whole cell & cell free assays to detect inhibitors of Aβ40
WB protocol to detect fecal Adipsin protein elevated in Notch gut toxicity
Binding assays to monitor tissue drug levels in brain tissue & plasma
Protein/antibody purification & characterization
Immunomagnetic positive selection of CD14 cells from human PBMCs
Recombinant protein expression & purification by metal chromatography
BCA, Bradford protein assays, Nanodrop
Antibody biotinylation
Dendritic cell antibody purification by protein A/G chromatography
SDS-PAGE, 2D gels, Western Blotting
ELISA, ELISpot
Cell/molecular biology
Primary & continuous cell line culture; transfections
Isolation & characterization of human T, B, DC cells
RNA/DNA isolation, subcloning, transformation, plasmid prep, PCR, restriction analysis, DNA electrophoresis, site-directed mutagenesis, primer design
Computer/software
Excel/PowerPoint, GraphPad Prism, Biostatistics
EDUCATION:
M.S. 1987 University of Texas, M.D. Anderson Cancer Center, Houston, TX
Dissertation: Evidence for Shared Epitopes between Intracellular and Extracellular Human B Cell Growth Factor.
B.S. 1977 New Mexico State University, Las Cruces, NM (Biology)
B.A. 1977 New Mexico State University, Las Cruces, NM (Chemistry)
RESEARCH EXPERIENCE:
2014-present Research Associate
University of Rhode Island
Institute for Immunology & Informatics, Viral Lab
Providence, RI
Dendritic cells were isolated from human PBMCs in order to look at the effects of viral infection. Immunomagnetic isolation of CD14 positive cells was used as a source of dendritic cells. A cytokine secreting cell line, HEK-293T, produced recombinant IL4 & GM-CSF used for the differentiation of CD14 positive cells into dendritic cells. Analysis of the viral effects on the DCs was done by Western Blot, Flow cytometry, Luminex, and RNA isolation. Mitochondrial fractionation of the DCs was also performed for analysis of viral effects.
Techniques: cell culture, Immunomagnetic isolation, Western Blot, Flow cytometry, mitochondrial isolation, transformation, isolation of plasmid DNA with restriction analysis, IL4 & GM-CSF ELISAs
2012-2014 Research Associate
University of Rhode Island
Institute for Immunology & Informatics, Immunology Lab
Providence, RI
Expression and purification of recombinant proteins for development of a vaccine delivery vehicle. Developed binding assay for mouse/human dendritic cell antibodies. Immunomagnetic isolation of dendritic and T4 cells from human PBMCs for coculture experiments with flow cytometric analysis of surface/intracellular markers looking for effects from the dendritic cell antibodies on the immune response.
Techniques: cell culture, transfections; purification of recombinant antibodies by Protein A/G on an Akta Avant; transformation, bacterial growth/induction with purification by metal chromatography; SDS-PAGE, Western Blot; plasmid preps, primer design, site-directed mutagenesis; endotoxin analysis/removal; Flow cytometry
2010-2012 Senior Research Associate
Molecular & Cell Biology, Protein Biochemistry
EpiVax, Providence, RI
Established molecular protocols for subcloning
Maintained cells for transfections & recombinant protein production
Chemical conjugation (Solulink) of anti-DEC to Tregitopes, HSA
Developed an indirect ELISA for a complement protein
Improved techniques for gel electrophoresis & western blotting
Performed binding studies using cell lines & FACS analysis
Monitored T cell activity & proliferation by ELISpot analysis
Performed MHC binding assays with analysis by time resolved fluorescence
Techniques: PCR, plasmid prep, DNA restriction analysis, transformation, subcloning, DNA electrophoresis, site-directed mutagenesis, primer design, cell culture, antibody chemical conjugation, ELISA, ELISpot, endotoxin testing, protein assays, SDS-PAGE, Western Blot, protein production
2009 Senior Research Assistant
Molecular & Cell Biology, Autism
Brown University, Providence, RI
Set up new research lab and adapted molecular/cell biology & protein biochemical methods to explore the unique genetics of autism
Techniques: cell culture, transformation, plasmid prep, transfection, restriction enzyme analysis, DNA gel analysis, SDS-PAGE, Western blot, LiCor, Nanodrop
2008-2009 Scientist
Neuroscience, Translational Biomarkers
Pfizer Research & Development
Established assay to detect multiple inflammatory proteins simultaneously using Luminex xMAP technology to help identify possible biomarker for neurodegeneration.
Developed protocols for examining cytokines in brain extractions and plasma to help identify a translatable transgenic mouse model of neuroinflammation.
Learned biostatistics/ANOVA and applied to analysis for significant trends in cytokine data.
Became skilled at autoradiography to apply to the development of a brain slice binding occupancy assay.
Supported the Gamma Secretase Modulator project by immunophenotyping peripheral blood B lymphocytes by FACS analysis to detect Notch toxicity.
Techniques: Luminex xMAP, brain extraction, biostatistics, autoradiography, brain sectioning, phosphorimaging, flow cytometry.
1997-2008 Scientist
CNS, Alzheimer’s disease
Pfizer Research & Development
Helped refine a fluorescent assay to detect inhibitors of b-amyloid protein aggregation.
Western blot analysis of transgenic mice brain homogenates, and pharmacology of APP processing after treatment with gamma secretase inhibitors.
Helped set up metabolic labeling/IP of cells for detection of inhibition of products of APP processing.
Developed in vitro whole cell and cell free assays with Aβ40 ELISA to measure inhibitors of β-amyloid production. Ran both primary screens for gamma secretase inhibitor project.
Set up Aurora-based gamma secretase reporter assay to detect inhibitors with selectivity between APP and Notch.
Developed Western blot protocol to detect fecal Adipsin protein which was elevated due to Notch-related gut toxicity.
Developed ex vivo and in vitro binding assays to monitor drug levels in brain and plasma
Performed routine antibody characterizations.
Adapted a technique to permit analysis of peripheral blood lymphocytes from mice, rats, dogs, and guinea pigs treated with gamma secretase inhibitors using whole blood and four-color immunofluorescence staining with flow cytometry, looking for a safety biomarker.
Helped develop and run cell free primary screen for beta secretase, increasing throughput by conversion from 96 to 384 well format.
Techniques: fluorimetry, cell culture, SDS-PAGE, Western Blot, ELISA, enzyme kinetics, membrane extraction, densitometry, metabolic labeling, immunoprecipitation, flow cytometry.
1996-1997 Associate Scientist (TEMP)
Drug Safety Evaluation, Cell Toxicology
Pfizer Research & Development
Continued research on drug toxicology in liver cells under GMP conditions.
Liver hepatocytes were isolated, cultured, and used to analyze in vitro drug effects on cell viability and lipid accummulation.
Developed sensitive radiometric assay for analyzing drug effects on -oxidation of fatty acids in mitochondria isolated from liver and cultured hepatocytes.
Adapted fluorescent and nonfluorescent staining techniques for the analysis of drug effects on lipid accummulation in cultured hepatocytes.
Designed experiments to look at the effects of drugs on triglyceride egression from cultured hepatocytes.
Techniques: primary cell culture, rat/dog liver perfusion, Lipid cell staining, -oxidation, SDH automated analysis, fluorimetry, fluorescent microscopy; mitochondrial isolation.
1994-1996 Senior Research Assistant
Surgical Research, Wound Healing
Rhode Island Hospital
Explored the mechanism of nitric oxide (NO)-mediated apoptotic tumor cell death by activated macrophages.
Transfected a NO-sensitive murine cell line with a eukaryotic expression vector containing the human BCl-2 gene, and the stably transfected cell line showed protection against macrophage killing as well as apoptotic-inducing and NO-donating drugs.
Studied the mechanism of induction of nitric oxide synthase at the RNA and protein levels in wound-derived rat macrophages.
Helped develop a biochemical purification scheme for murine macrophage arginase, and design oligonucleotide probes to clone the gene.
Techniques: cell culture, retroviral and liposome-mediated transfection, immunofluorescence, macrophage cytoxicity/binding assays, 3H-thymidine release/uptake, 51Cr Release, MTT assay, SDS-PAGE, Western blot /densitometry, RNA/DNA isolation, RT-PCR, subcloning, transformation, plasmid purification, DNA labeling, S1 nuclease protection assay, Northern blot, autoradiography, DNA electrophoresis.
1992-1994 Pre-doctoral Fellow
Molecular Pathology
University of Texas, M.D. Anderson Hospital
Investigated the in vitro growth of Non-Hodgkin’s Lymphoma B cell lines (isolated from patients’ blood). A human B cell growth factor (IL-14), expressed only in high grade NHL-B's and activated normal T cells but not in normal B cells, was proposed to be an autocrine growth factor .
IL-14 was subcloned into a eukaryotic gene expression vector, transfected into Cos cells, and the expressed recombinant protein as well as IL-14 isolated from NHL-B cell lines showed activity on target normal B cells.
Antisense oligomers directed against IL-14 were found to inhibit NHL-B proliferation in a dose-dependent manner.
Studied the proposed autocrine growth loop was using drugs known to interfere with signal transduction (CsA, FK506, Rapamycin), protein trafficking (monensin), growth factor/receptor interactions (suramin), and cell differentiation (retinoic acid).
Examined the induction of apoptosis in NHL-B cells following drug treatment.
Techniques: cell culture, DNA fragmentation (Burton) assay, DNA ladder electrophoresis, Tunel, immunohistochemistry, EM, flow cytometry, cDNA library construction/Ab screening, isolation of DNA/RNA, DEAE-Dextran transfection, subcloning, transformation, random prime labeling, Northern/Southern blot, RT-PCR, immunoprecipitation, column chromatography, 3H-thymidine uptake, MTT assay.
1988-1992 Associate Scientist
Infectious Diseases
Pfizer Research & Development
Purified and characterized mammalian topoisomerase, bacterial topoisomerase, urease and ATPase.
Developed a novel purification scheme for DNA gyrase from quinolone-susceptible and resistant clinical isolates of Campylobacter jejuni.
Developed a cell colony-forming assay to examine the effects of quinolones on the survival of parental and drug-resistant CHO cell lines.
Developed a microplate fluorescent cytotoxicity assay for the evaluation of anti-cancer drugs on a mouse lymphoma cell line.
Analyzed bacterial ATPase was as a possible target for anti-ulcer therapy.
Techniques: column chromatography, glycerol gradient fractionation, DNA relaxation/catenation assays, SDS-PAGE, subunit complementation studies, flow cytometry, membrane extraction, spectrophotometric kinetic and phosphate assays, generation of inverted vesicles, FPLC, cell culture, fluorescent assay.
1987-1988 Senior Research Assistant
Pediatric Gastroenterology
Rhode Island Hospital
Developed and characterized long-term cultures of mammalian visceral smooth muscle cells to replace freshly isolated cells in experimental protocols.
Isolated smooth muscle cells by enzymatic digestion of gastrointestinal tissues.
Compared contraction and relaxation responses to calcium and/or neuropeptides of freshly isolated cells versus long-term culture cells. Cells were also analyzed for phosphoinositides
.
Techniques: primary/continuous cell culture, phase-contrast microscopy, SDS-PAGE, Western blot, immunofluorescence.
1985-1987 M.S. graduate student
Pathobiology
University of Texas, M.D. Anderson Hospital
Purification and characterization of a putative high molecular weight intracellular precursor for human B cell growth factor (BCGF).
Helped generate heterotypic and monoclonal antibodies to the homogeneously pure intracellular BCGF (IC-BCGF).
Constructed affinity columns using purified IC-BCGF antibodies to one-step purify both intracellular and extracellular BCGF (EC-BCGF).
Analyzed purified high (IC) and low (EC) weight BCGF for homologous bioactivities and characteristics.
Techniques: ion-exchange/molecular sieve/affinity chromatography, isoelectric focusing, SDS-PAGE, silver stain, Western blot, mRNA isolation, 3H-thymidine uptake, ELISA, Ouchterlony RID, cellular fractionation, cell culture, monoclonal antibody production, small animal surgery.
1983-1985 Senior Research Assistant
Pathobiology
M.D. Anderson Hospital & Cancer Center
Purified and characterized a putative intracellular precursor for human IL-1.
Characterized long-term human B/T lymphocytes.
Analyzed patient joint fluid samples for IL-1.
Techniques: mouse thymocyte IL-1 assay, 3H-thymidine uptake, cell culture, column chromatography, isoelectric focusing, SDS-PAGE, small animal surgery.
1981-1983 Research Assistant
Urology Research
M.D. Anderson Hospital & Cancer Center
Developed 2-dimensional gel analysis system for tissue, plasma, and urine samples
Analyzed patient samples in search of a protein marker specific for adenocarcinoma of the prostate.
Techniques: column chromatography, 2-dimensional gel electrophoresis, silver staining, computer digital analysis.
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