Research Associate

PRANAL J DAKLE

**** **** **** ******, ******.*****@*****.***

Los Angeles, CA-90007 Phone: 617-***-****

SUMMARY OF SKILLS

Techniques:

• Formulation of liposomes, micelles, tablets, capsules, suspensions, emulsions, lotions, ointments

• Flow Cytometry / FACS Analysis -BD FACSCalibur, FACScan

• Tissue Culture and aseptic technique

• Cell Based Assays and Immunoassays

• Molecular Biology Techniques: SDS-PAGE, Western Blot, PCR, Gel Electrophoresis, Plasmid Preparation, DNA &RNA preparation (Isolation and Purification), Transfection, cDNA synthesis

• Biochemical techniques like ELISA, BCA Protein Assay

• Chromatography methods like HPLC, Paper, Column, Thin layer chromatography

• Animal handling, injection (IV and IP) and dissection

• Microscopy techniques like Fluorescence microscopy- Immunofluorescence

• Particle size analysis by Coulter Counter and Zeta Potential Analysis

• Spectroscopy technique - UV/Vis spectroscopy (Jasco)

• Knowledge of GLP and GMP regulations

Instrumentation:

FACS, UV-VIS Spectrophotometer, HPLC, Nikon Eclipse E400 microscope, Plate reader, Rotary evaporator, Bioquant Microscope, Coulter Counter Particle Size Analyzer, Zeta Potential Analyzer, Probe Sonicator, pH meter, Freeze dryer, Vi-Cell, EVOS Fluorescence Microscope

EDUCATION

Northeastern University Boston, MA

Master of Science in Pharmaceutical Sciences Aug 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

Pune University Pune, India

Bachelor of Pharmacy in Pharmaceutical Sciences July 2005-May 2009

Coursework: Pharmaceutics, Pharmacology, Biotechnology, Microbiology, Biochemistry, Pharmaceutical Analysis, Biopharmaceutics and Pharmacokinetics, Pharmacognosy

RELATED WORK EXPERIENCE

Agensys Astellas Los Angeles, CA

Research Associate June 2011- Sept 2011

Oncology Project: Antibody Drug Conjugate Biology

• Performed wide range of cell based cytotoxicity assays on various anticancer Spirogen compounds

• FACS-based assays to measure protein expression on the surface of mammalian cells

• Studied Apoptosis/Cell death assays by flow cytometry with the help of Annexin V

• Performed MDR1 shift assay using flow cytometry

• Separated protein extracts from cell lysates by SDS-PAGE and detected level of protein using Western Blot

• Extensive mammalian cell culture

• Immunofluorescence(IFA) assays to detect the surface expression of various antibodies in different cell lines

Dr. Vladimir Torchilin’s Lab Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University Boston, MA

Graduate Research Associate April 2010- June2011

Advisor: Dr. Erez Koren

Oncology Project: 1. Multifunctional pH sensitive nanocarriers for cancer treatment

• Formulated micelles containing pH sensitive bonds and PEGylated TAT peptide for uptake by tumor cells

• Synthesized and purified PEG2000-hydrazone-PE

• Freeze drying/ lyophilization

• Maintained mammalian cancer cell lines (B16F10, Hela) and adept in cell culturing under aseptic conditions

• Fluorescence Microscopy based assays showed fluorescence indicating internalization of pH sensitive micelles

• Studied cell interaction studies of micelles with tumor cell lines using Flow Cytometry (FACS analysis)

• Carried out RP-HPLC based assays to determine the drug loading capacity and also stability of formulation

• 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

• Performed in invivo studies on C57BL6 tumor bearing mice by tail vein injection

• Dissection of mice and tumor collection

• TUNEL assay and Immunohistochemistry

Oncology Project 2. Multifunctional Immunoliposomes for tumor targeting

• Formulated immunoliposomes containing pH sensitive bonds, mAb 2C5 and PEGylated TAT peptide for uptake by tumor cells

• Tissue Culture and aseptic technique

• Determined the stability of the formulation by using RP-HPLC based assays

• Performed ELISA and compared the activity of formulation with standard 2C5 and IgG antibodies

• Immunofluorescence Microscopy based assays showed internalization of pH sensitive liposomes

• Studied cell interaction studies of immunoliposomes with tumor and normal cell lines using Flow Cytometry (FACS)

• ADCC (Antibody Dependent Cell Cytotoxicity)

• Carried out cell viability studies by performing cell based cytotoxicity assays

Department of Pharmaceutical Sciences,Northeastern University Jan 2010 – April 2010

Research Associate:

• Cell Culture Techniques- Cell Line Initiation, Splitting and Passaging

• FACS-based assays to measure protein expression on the surface of mammalian cells

• Plasmid Preparation, DNA &RNA preparation (Isolation and Purification), Transfection

• Plasmid Preparation from E.coli and its purification

• Protein Expression Studies- SDS-PAGE, Western Blot, PCR, Gel Electrophoresis, Agarose Gel Electrophoresis, IEF

• Bioanalytical techniques- ELISA, BCA Protein Assay, Enzyme Kinetics Assays

• Separated protein extracts from mammalian cell culture by SDS-PAGE and detected B-Actin protein using Western Blot using Bio-Rad Mini-Gel Box Electrotransfer

• Performed transfection of NIH3T3 cells by pEGFP (Green Fluorescence Protein) followed by ELISA to quantify the amount of EGFP in the same

• Performed PCR and amplified cDNA libraries via RT-PCR using Go Taq DNA Polymerase; Promega

• Immunohistochemistry

Gripil Pharmaceuticals Pvt, Ltd Mumbai, India

Intern Nov 2007-Dec 2007

• Assisted in manufacturing processes of creams, ointments and tablets

• Performed Quality Control tests for tablets, like Disintegration, Dissolution, Hardness and Friability tests

Pune University – Lab Courses July 2005-May 2009

• Pharmaceutics: Formulated tablets, capsules, suspensions, solutions, emulsions, gels, ointments, parenterals

• Chemistry: UV/Visible spectrometry, IR spectroscopy, chromatography (HPLC, TLC), titrimetry

• Pharmacology: Trained on animal handling techniques for mice and rats, drug administration through I.V, I.M, S.C, I.P and oral routes

COMPUTER KNOWLEDGE

• Skilled in using Macintosh and Windows

• Microsoft word, Excel, Power Point, Adobe Photoshop

• Other software- FCS Express, Cell Quest Pro, Graph Pad Prism, Image J, Image Lab

References would be furnished upon request

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