Medical System

Location:

United States

Posted:

July 30, 2012

Contact this candidate

Resume:

Hitesh K. Jindal, Ph.D.

** ***** **** **** • West Roxbury, MA 02132

Tel: 617-***-**** (Home) • 617-***-**** (Cell) • E-mail: ********@*****.***

SUMMARY:

Ph.D. (Biochemistry). Scientist with extensive experience in protein biochemistry and cell biology

Experience includes:

• Extensive hands-on experience in purification of native and recombinant proteins, characterization of purified proteins, generation and purification of poly clonal and monoclonal antibodies, enzymology, quantitative proteomic analysis, phosphoproteome analysis, post-translational modifications (phosphorylation and palmitoylation), bioconjugation/modification of proteins, protein-protein interactions, assay development, immunoassays, immunofluorescence, cell culture and generation of stable cell lines.

• Areas of expertise in ion channels research, signal transduction, in-vitro ADMET, and ligand binding assays.

• Ability to direct, perform, analyze, and interpret basic research projects involving protein biochemistry.

• Extensive hands-on experience in managing biochemistry laboratories and ability to train and mentor junior scientist, graduate students and technicians.

TECHNICAL SKILLS:

Protein Chemistry:

• Purification of native and recombinant proteins

• Expertise in the use of FPLC, and other conventional chromatographic techniques including gel filtration, ion exchange, hydrophobic interactions and affinity chromatography

• Characterization of native and recombinant purified proteins for their biological functions

• Enzymology-enzymatic assays and their design using radio isotopes, UV/visible and fluorescence spectroscopy

• Sample preparation for isoelectric focusing (IEF) and 2-Dimensional differential in gel electrophoresis

(2-D DIGE)

• Electrophoretic techniques including native and SDS-PAGE, IEF, 2 D-PAGE, 2 D-DIGE, and Western blotting.

• Protein-protein interaction/binding studies employing co-precipitation, co- immunoprecipitation, blot overlay, pull down assays and co-transfection methods

• Photo affinity and covalent affinity labeling of proteins with Cy dyes, NHS biotin and NHS fluorescein

• Partial tryptic digestion, amino acid composition and amino acid sequence analysis

• In vitro phosphorylation of purified proteins using multiple kinases, metabolic radio labeling of proteins in mammalian cells and Sf-9 cells, phosphoamino acid analysis, and identification of phosphorylation sites employing 2-dimensional phosphopeptide mapping of tryptic peptides

• Reduction, carboxy methylation, in-gel trypsin digestion of proteins and extraction of trypsin digested peptides for LC-MS/MS analysis

• Enrichment of phosphopeptides using TiO2 affinity resin

• Radioactive, fluorescence, and spectrophotometric enzyme/metabolic assay, enzyme kinetics, automated liquid handling, and TLC

Immunochemistry:

• Production of polyclonal antibodies against purified native and recombinant proteins and their affinity purification.

• Production of ascites, purification of monoclonal antibodies and their use in ELISA, Immunoblotting, immunoprecipitation, immunofluorescence and subcellular localization of antigens in cells and tissue samples

Cell Biology:

• Tissue culture (mammalian cells and insect cells)

• Cell migration/invasion assay

• Operation of fluorescent microscope and immuno-fluorescence microscopic studies to detect native as well as recombinant proteins expressed in mammalian cells and Sf-9 cells

PROFESSIONAL EXPERIENCE:

2012. -, Boston Bioproducts, Ashland, MA, Senior Scientist-Protein Biochemistry

• Expression and purification of recombinant proteins

• Expression and purification of antibodies

• Bio-conjugation of purified proteins and antibodies

2005 – Dec. 2011, Alpert Medical School of Brown University, Cardiovascular Research Center, Rhode Island Hospital, Providence, RI, Assistant Professor (Research)

• Characterized the effect of C-terminal palmitoylation on expression, turnover and trafficking of voltage-gated potassium channel Kv1.5.

• Generated stable cell lines over expressing wild type and palmitoylation mutant forms of voltage-gated potassium channel Kv1.5 for electrophysiological studies and for studying trafficking of cell surface channel proteins.

• Purified affinity-tagged recombinant voltage-gated potassium channel KvLQT1 and HERG proteins expressed in transgenic rabbit hearts using affinity chromatography and identified their binding partners by mass spectrometry.

• Established proteomic 2-Dimension Fluorescence Differential In-Gel Electrophoresis (2-D DIGE) system in conjunction with LC-MS/MS analysis to elucidate differential protein expression in transgenic rabbits hearts generated as human models for long QT syndrome 1 (LQT1) and long QT syndrome 2 (LQT2).

• Optimized conditions, standardized protocols and trained undergraduate students for effective use of 2-D DIGE system.

• Validated results of 2-D DIGE and LC-MS/MS analysis in LQTS rabbit hearts, which revealed differential upregulation of the expression and activities of the key ATP-generating enzymes, suggesting metabolic remodeling in these transgenic rabbit hearts.

• Performed enrichment of phosphopeptides to demonstrate the effect of sex hormones on phosphoproteome in transgenic LQT2 rabbit hearts, and to elucidate the potential pro/anti-arrhythmic role of sex hormones.

2002- 2005, Tecan Boston, Medford, MA, Senior Scientist

Worked as an active member of the team adapting micro titer plate-based in vitro ADMET enzyme assays to an automated high-throughput low-volume microfluidic platform

• Managed the biochemistry, and tissue culture facility.

• Established laboratory protocols and studied the metabolic clearance of drugs using a centrifugally-driven, disposable microfluidic device (LabCDTM) with cryo-preserved human hepatocytes.

• Developed an assay system to determine multidrug resistance P-glycoprotein associated ATPase activity using a novel phosphate binding protein as a detection probe.

• Handled robotic liquid handling system as well as UV/visible and fluorescence spectroscopy.

2001- 2002, SatCon Technology Corporation, Cambridge, MA, Senior Biochemist

Worked as a Senior Biochemist in start-up of SatCon biosensing program directed at microbiological testing market with technical focus on the rapid detection and identification of biological targets

• Managed the biochemistry/microbiology and wet chemistry laboratory.

• Collaborated with the engineers to develop microfluidic devices, flow cell and photonics, and scientists in the area of microbiology, food sciences and surface chemistry.

• Demonstrated the specific capture of microbes and proteins on regenerable solid platform engineered with affinity probes on self-assembled polyethylene oxide monolayer.

1998 - 2001, Harvard Medical School, Beth Israel Deaconess Medical Center, Boston, MA, Instructor

• Characterized novel binding partner of Related Adhesion Focal Tyrosine Kinase (RAFTK), AER protein-product of the BRCA2 transcription unit in breast cancer.

• Expressed and purified recombinant AER protein expressed as GST-fusion protein in E.coli, developed polyclonal antibodies against purified AER fusion protein, and generated stable cell lines expressing AER protein.

• Demonstrated the in vivo and in vitro association and in vitro co-localization of AER and RAFTK proteins and co-localization of AER protein with clathrin using confocal immunofluorescence microscopy suggesting a likely biological role of AER protein in vesicular trafficking

1994 - 1998, Tufts University School of Medicine, St. Elizabeth’s Medical Center, Boston, MA, Research Associate

• Purified a novel membrane bound Neurofibromatosis type 2 (NF2) tumor suppressor protein from human erythrocytes to homogeneity using FPLC and other conventional chromatographic techniques including gel filtration, ion exchange chromatography. Characterized the biological functions of the purified NF2 protein.

• Standardized various protein kinase assays (i.e. PKC, cAMP-dependent kinase, and casein kinase type I and II), optimised conditions for the in vitro phosphorylation of the cytoskeletal membrane proteins and demonstrated the specific loss of protein kinase activities in senescent erythrocytes.

1990 - 1994, University of British Columbia, Dept. of Biochemistry, Vancouver, Canada, Research Associate

• Purified nonstructural protein (NS-1) of minute virus of mice expressed in Sf-9 using an immuno-affinity purification strategy.

• Established enzymatic assays and developed standard operating protocols for covalent and photo-affinity labeling to characterize the biological functions of the purified NS-1 protein, as ATP dependent helicase enzyme.

• Elucidated the post-translational modification of NS-1 protein by the in vitro phosphorylation of purified protein by multiple kinases, metabolic labeling of protein, phosphoamino acid analysis, and identified two potential phosphorylation sites on NS-1 protein employing 2-dimensional phosphopeptide mapping.

EDUCATION:

Ph.D., Biochemistry, Kurukshetra University (National Dairy Research Institute), Karnal, India.

• Purified two native enzymes involved in fatty acid synthesis (i.e. fatty acid synthetase and acetyl-CoA carboxylase) from goat mammary gland using conventional chromatographic techniques including gel filtration and ion exchange chromatography.

• Established and maintained a tissue culture lab.

• Standardized enzymatic assays to elucidate the effect of hormones on the activities and rate of synthesis of enzymes involved in fatty acid synthesis and NADPH+ generating enzymes.

M.S., Biochemistry, Kurukshetra University (National Dairy Research Institute), Karnal, India.

AWARDS:

• Junior Research Fellowship Award, Indian Council of Agricultural Research, India, 1981-1983

• Senior Research Fellowship Award, Indian Council of Agricultural Research, India, 1983-1986

• Milheim Foundation Award for Cancer Research, July 2000-June 2001

COMPUTER SKILLS:

• Proficient in Microsoft Office (Microsoft Word, Excel, and Power point), Adobe Acrobat, 2-D DIGE analysis software (ImageQuantTL and DeCyder), and Image J.

• Proficient in electronic record keeping

TRAINING/WORKSHOPS:

• Agricultural Research and Management Training: National Academy for Agricultural Research and Management, Hyderabad, India. (December 1987-May 1988)

• Genomics and Proteomics Hands-on Workshop: From Sample Preparation to Data Analysis, National Jewish Health Center and University of Colorado, Denver, Colorado. (July 8-17, 2009)

PUBLICATIONS:

Peer-Reviewed Original Articles and Book Chapters:

1. Jindal, H.K., Merchant, E., Balschi, JA, Zhangand, Y and Koren, G. (2012) Proteomic analyses of transgenic LQT1 and LQT2 rabbit hearts elucidate increase in expression and activities of energy producing enzymes. J. Proteomics (In Press)

2. Beleych, A.E., Sanson, S.E., Terentyeva, R., Ho, H.T., Nishijima, Y., Martin, M.M., Jindal, H.K., Rochira, J.A., Kunitomo, J.A., Abdellatif, M., Elton, T.S., Gyorke, S. and Terentyev, D. (2011) MicroRNA -1 and -133 increase arrythmogenesis in heart failure by dissociating phosphatase activity from RyR2 complex. PLoS One, 6 (12): e28324

3. Ren, X-Q, Liu, G-X, Organ-Darling, L.E., Zheng R, Roder, K., Jindal, H.K., Centracchio, J., McDonald T.V., Koren, G. (2010) Pore mutants of HERG and KvLQT1 downregulate the reciprocal currents in stable cell lines. Am. J. Physiol. Heart Circ. Physiol. 299: H1525-H1534.

4. Jindal, H.K., Folco, E., Liu, G.X., and Koren, G. (2008) Post-translational modification of voltage-dependent potassium channel Kv1.5: COOH-terminal palmitoylation modulates its biological properties. Am. J. Physiol. Heart Circ. Physiol. 294: H2012-21.

5. Jindal, H. K., Yoshinaga, K., Seo, P-S., Lutchman, M., Dion, P.A., Rouleau, G.A., Hanada, T. and Chishti, A. H. (2006) Purification of the neurofibromatosis type 2 (NF2) tumor suppressor protein from human erythrocytes. Can. J. Neurol. Sci. 33: 394-402

6. Jannatipour, M., Dion, P., Khan, S., Jindal, H. K., Fan, X., Laganiere, J., Chishiti, A.H., and Rouleau, G.A. (2001) Schwannomin isoform-1 interacts with syntenin PDZ domains. J. Biol Chem. 276: 33093- 33100.

7. Peters, L. L., Jindal, H. K., Gwynn, B., Korsgren, C., John, K. M., Lux, S. E., Mohandas, N., Cohen, C. M., and Brugnara, C. (1999) Mild spherocytosis and altered red cell ion transport in protein 4.2-null mice. J. Clin. Invest. 103: 1527-1537

8. Chishti, A. H., Kim, A. C., Marfatia, S. M., Lutchman, M., Hanspal, M., Jindal, H. K., et al. (1998) The FERM domain: A unique module involved in the linkage of cytoplasmic proteins to the membrane. Trends Biochem. Soc. 23: 281-282

9. Jindal, H. K. and Cohen, C. M. (1996) Identification and down regulation of protein kinase C isozymes in human blood cells. Mol. Biol. Cell. 7: 521

10. Astell,C. R., Liu, Q., Harris, C. E., Brunstein, J., Jindal, H. K. and Tam, P. Minute Virus of Mice cis-acting sequence required for genome replication and the role of the transacting viral protein, NS-1 in W. Cohn and K. Moldave (eds). Progress in Biochemistry and Molecular Biology, Academic Press, 1996, Vol. 55, p 245.

11. Jindal, H. K., Ai, Z., Gascard, P., Horton, C. and Cohen, C. M. (1996) Specific loss of protein kinase activities in senescent erythrocytes. Blood, 88: 1479-1487.

12. Jindal, H. K., Yong, C. B., Wilson, G. M., Tam, P. and Astell, C. R. (1994) Point mutations in the NTP-binding motif of MVM NS-1 protein uncouple ATPase and DNA helicase functions. J. Biol. Chem. 269: 3283-3289.

13. Vishwanatha, J.K. Jindal, H. K., and Davis, R. G. (1992). The role of primer recognition proteins in DNA replication: Association with nuclear matrix in HeLa cells. J.Cell Sci. 101: 25-34.

14. Wilson, G. M., Jindal, H. K., Yeung, D. E., Chen. W. and Astell, C.R. (1991) Expression of minute virus of mice major non-structural protein in insect cells: purification and identification of ATPase and helicase activities. Virology, 185: 90-98.

15. Jindal, H. K., Chaney, W. G., Anderson, C. A., Davis, R. G. and Vishwanatha, J. K. (1991) The protein tyrosine kinase substrate, calpactin 1 heavy chain (p36), is a part of the primer recognition protein complex that stimulates DNA polymerase a activity. J.Biol.Chem. 266: 5169-5176.

16. Jindal, H. K., Anderson, C. A., Davis, R. G. and Vishwanatha, J. K. (1990) Suramin affects DNA synthesis in HeLa cells by inhibition of DNA polymerases. Cancer Research 50: 7754-7757.

17. Jindal, H. K. and Vishwanatha, J. K. (1990) Functional identity of a primer- recognition protein as phosphoglycerate kinase. J. Biol. Chem. 265: 6540-6543.

18. Jindal, H. K. and Vishwanatha, J. K. (1990). Primer-recognition proteins in HeLa cells: purification and partial characterization. Biochemistry 29: 4767-4773.

19. Jindal, H. K. and Panday, R. S. (1988) Studies on the regulation of enzymes related to fatty acid synthesis in goat mammary glands using explant culture system. Indian J.Biochem. Biophys. 25: 428-433.

20. Jindal, H. K., Bansal, V.S., Kasinathan, C., Larroya, S. and Khuller, G. K. (1983). Effect of carbon source on the polar fatty acids of Microsporum gypseum grown at different temperatures. Experientia 39: 151-153.

RECENT ABSTRACTS/PRESENTATIONS:

1. Rieder, R., Jindal, H.K., Laibinis, P., and Weigle, B. (2002). An ultra-senstive optical biosensor detection of single microbes.

2. Rieder, R., Howatt, J., Jindal, H.K., Pavarnic, P. and Senecal, A. (2002). A Microbiological Lab-on-a-chip.

3. Ommert, S., Towel, T., Haspel, H., Jindal, H.K., Lance, G. and Bansal, P. (2003). ADMET assay on Tecan’s LabCD-ADMET system: MDR-1/P-glycoprotein (Pgp) ATPase assay: Association for Laboratory Automation.

4. Cai, H., Smith, D., Reddy, A., Johnson, K., Winter, R., Blanchette, J., Jindal, H.K., Schmid, N., Bansal, P.,

Kellogg, G., Narayanan, S. (2003). A fisibility study of LabCD format for the prediction of human hepatic clearance using cryopreserved human hepatocytes. ISSX, 12th North American Meeting, Providence, Oct.12-16.

5. Gleich, L., Karr, C., Towle, T., Hickey, M., Jindal, H.K, Schilling, E., Carvalho, B., Van Delinder, G.,

Meredith, J., Johnson, B., Whitney, S., Bansal, P. and Contarino, M. (2004) LabCD-ADMET System Validation For Serum Protein Binding Assays. SBS 10th Anniversary Conference and Exhibition, Orlando, Florida, Sep. 11-15

6. Gleich, L., Towle, T., Shea, M., Ommert, S Hickey, M., Jindal, H. K, Schilling, E., Carvalho, B., Van Delinder, G., Meredith, J., Johnson, B., Whitney, S., Contarino, M. and Bansal, P. (2004) LabCD-ADMET System Validation For Cytochrome P450 Inhibition Assays. SBS 10th Anniversary Conference and Exhibition, Orlando, Florida, Sep. 11-15

7. Jindal, H. K. and Gideon Koren, (2009) Proteomic analysis of KvLQT1 and HERG –associated proteins. Biophysical Society, 53rd Annual Meeting, Boston, MA, February 28- March 4.

8. Jindal, H. K. and Gideon Koren, (2009) Determination of KvLQT1 and HERG interacting proteins using 2-D DIGE and proteomics studies. 30th Annual Scientific Sessions of the Heart Rhythm Society, Boston MA, May 13-16.

REFERENCES:

Dr. Gideon Koren, MD

Professor of Medicine

Director, Cardiovascular Research Center,

Cardiology Division

Rhode Island Hospital

Alpert Medical School of Brown University

Tel: 401-***-****

E mail:******@********.*** or ************@*****.***

Dr. Ulrike Mende, MD

Associate Professor of Medicine

Cardiovascular Research Center,