Medical Analysis
Resume:
P RANAL J DAKLE
p *****.*****@*****.***
Boston, MA-02215 Phone: 617
SUM MARY OF SK I L LS
Techniques:
• Flow Cytometry (Multi color FACS analysis) -BD FACSCalibur, BD FACScan, BD FACSCanto I I and Flow Jo for
analysis of data
• Cell Culturing (Mammalian Cell Culturing)
• Cell Based Assays
• Immunoassays: L uminex Based Assays, ELISA, ELISPOT
• Molecular Biology Techniques: SDS-PAGE, Western Blot, Real-time PCR, Gel Electrophoresis, P lasmid
P reparation, DNA &RNA preparation (Isolation and Purification), Transfections
• Microscopy techniques- Immunofluorescence, Immunohistochemistry, TUNEL staining
• Animal handling: t ail tip for genotyping, non-surgical collection of body f luids, euthanasia and t issue harvest,
i njections, monitoring for experimental endpoints, isolation of bone marrow, dissection to isolate organs like
spleen, uterus, placenta, lymph nodes etc
• Particle size analysis by Coulter Counter and Zeta Potential Analysis
• Spectroscopy technique - UV/Vis spectroscopy (Jasco)
• Formulation of l iposomes, micelles, tablets, capsules, suspensions, emulsions, lotions, ointments
• Ordering lab supplies, Inventory and Budget management
• Knowledge of GLP and GMP regulations
Instrumentation:
FACS, 7300 Real-Time PCR System, ChemiDoc, UV-VIS Spectrophotometer, N ikon Eclipse E400 microscope, P late reader,
Rotary evaporator, Bioquant Microscope, Coulter Counter Particle Size Analyzer, Zeta Potential Analyzer, Probe
Sonicator, pH meter, Balance, F reeze dryer, Vi-Cell, EVOS Fluorescence Microscope
WORK EXPER I E NCE
Child ren’s Hospital Boston/ Ha rva rd Medical School/ Brigham & Women’s Hospital
Boston, MA
T ransplantation Research Center ( Immunology Based Lab)
J an 2012- P resent
• Performed Intra-Cellular Cytokine Staining, Int racellular FoxP3 Staining of Splenocytes/Lymphocytes/Uterine
cells and analyzed results by multi color Flow Cytometry
• Studied Protein Expression by SDS-PAGE, Western Blot, PCR, Flow Cytometry
• Analyzed cell culture supernatants by ELISA, Luminex, ELISPOT to determine the concentration of different
cytokines
• Isolated and stained mouse splenocytes
• Studied Apoptosis induction by Dexamethasone/Hydrogen Peroxide confirmed by Annexin V and 7-AAD
• Performed and analyzed Phagocytosis assays by Flow Cytometry and Inverted Microscopic analysis
• Isolated subsets of cells by magnetic and f low based cell sorting
• Generated Tregs [CD4+CD25+Foxp3+cells] ex vivo f rom CD25- naive CD4 T cells
• Executed Mixed Lymphocyte Reaction (MLR) with responders and i rradiated stimulators (cesium source 3000rads)
• Animal handling: t ail tip for DNA genotyping, non-surgical collection of body f luids, euthanasia and tissue harvest,
i njections, monitoring for experimental endpoints,Isolation of Bone Marrow, dissection to isolate organs like
spleen, uterus, placenta, lymph nodes etc
• Isolated RNA from mice organs and cells and performed Real-Time PCR using SYBR green and TaqMan probes
• Immunohistochemistry of frozen and paraffin embedded sections
• Immunofluorescence staining to detect the expression of antibodies in cell lines
• Performed different cell based assays using Alamar Blue
• Managed ordering of lab supplies, Inventory and Budget
• Publication: Co-author for “TIM-3 Regulates Innate Immune Cells to Induce Fetomaternal Tolerance” published in
T he Journal of Immunology
Co-author for “ B7h (ICOS L) maintains tolerance at the fetomaternal interface” accepted for publication in The
A merican Journal
of Pathology
T ufts Medical Center ( Molecula r Oncology Research I nstitute)
Boston, MA
Oncology P roject: Antibody Discovery and Biomarker Study
O ct 2011-Dec 2011
• Performed SDS-PAGE and Western Blot analysis to detect the presence of MMP14 in different samples
• Studied Tube Formation, Migration and Invasion by cells in presence of different Antibody supernatants
• Performed ELISA to detect the presence and level of different Biomarkers for Cancer study in patient samples
using MSD (Meso Scale Discovery)
• Isolated RNA from patient samples and performed Real-Time PCR
• Detected the expression of MMP-14 Antibody in different cell lines by Flow based assays
Agensys Astellas
L os Angeles, CA
Research Associate J une 2011- Sept
2011
Oncology P roject: Antibody D rug Conjugate Biology
• Performed wide range of cell based cytotoxicity assays on various anticancer Spirogen compounds
• FACS-based assays to measure cellular proliferation and protein expression on the surface of mammalian cells
• Tested monoclonal Antibodies on a panel of different Cancer Cell Lines
• Studied Apoptosis/Cell death assays by f low cytometry using Annexin V staining
• Performed MDR1 shift assay using f low cytometry
• Estimated protein concentration of cell lysate by spectrophotometry
• Separated protein extracts from cell lysates by SDS-PAGE and quantified protein expression using Western
B lot
• Maintained and cryopreserved mammalian cell lines
• Immunofluorescence assays (IFA) and Immunocytochemistry assays to detect the surface expression of various
antibodies in different cell lines
Center for Pha rmaceutical Biotechnology and Nanomedicine, Northeastern University
Boston, MA
G raduate Research Associate J an
2010- June2011
• Formulated micelles containing pH sensitive bonds and PEGylated TAT peptide for uptake by tumor cells
• Formulated immunoliposomes containing pH sensitive bonds, monoclonal antibody 2C5 and PEGylated TAT
peptide for uptake by tumor cells
• Performed capillary electrophoresis to separate liposomes by their charge
• Studied internalization of pH sensitive micelles by Fluorescence microscopy
• Studied cell interaction of micelles with tumor cell lines using Flow Cytometry (FACS analysis)
• Characterized micelles for size and charge using Coulter Counter (N4 Plus) and Zeta potentiometer (BIC)
• Carried out cell viability studies by cell based cytotoxicity assays (MTT, CTG, Alamar Blue)
• Stained tissues for detection of apoptosis by TUNEL assay
• Performed ELISA to quantify protein in formulation
Northeastern University, Department of Pha rmaceutical Sciences
Boston, MA
Research Associate: J an 2010- April
2010
• Plasmid Preparation, DNA &RNA preparation (Isolation and Purification), Transfection
• Protein Expression Studies- SDS-PAGE, Western Blot, PCR, Gel Electrophoresis, Agarose Gel Electrophoresis
• Transfection of NI H3T3 cells by pEGFP (Green Fluorescence Protein)
E D UCAT IO N
• Northeastern University
Boston, MA
• Master of Science in Pharmaceutical Sciences A ug
2009-May 2011
• Specialization in Pharmaceutics and Drug Delivery Systems
Coursework
• Biochemistry, Molecular Cell Biology, Concepts in Pharmaceutical Sciences, Advanced Physical Pharmacy,
Pharmaceutical Sciences Laboratory, Advanced Drug Delivery Systems, Drug Design and Evaluation, Biometrics,
Pharmacokinetics and Drug Metabolism, Bioethics
P une University
P une, I ndia
• Bachelor of Pharmacy in Pharmaceutical Sciences J uly
2005-May 2009
• Coursework: Pharmaceutics, Pharmacology, Biotechnology, Microbiology, Biochemistry, Pharmaceutical Analysis,
B iopharmaceutics and Pharmacokinetics, Pharmacognosy
CO MP U TER KNOWLEDGE
• Skilled in using Macintosh and Windows
• Microsoft Word, Excel, Power Point, Outlook, Adobe Photoshop
• Other software- Flow Jo, FCS Express, FACS Diva, Cell Quest Pro, Graph Pad Prism, Softmax Pro, Image J,
I mage Lab
References would be furnished upon request
Contact this candidate