*** * ***** **** **, Chapel Hill, ***** 919-***-**** **********@*****.***
POLINA DANSHINA, Ph.D.
Professional Profile
Biochemist (enzymologist) with 15 years of proven experience in laboratory research on cell metabolism, fermentation, drug discovery, HTS
Reproductive biologist with expertise in Assisted Reproductive Technology
Highly motivated and organized with success in working both independently and in a team
Skills
Biochemistry: Recombinant protein expression and purification,Chromatography LC-MS, HPLC, GC, IC, ICP, UV-Vis, Karl Fisher, Enzyme kinetics, Activity assays, Immunochemistry, High throughput screening
Cell biology: Primary cell culture, cyto and histochemistry, cryotomy and sectioning, cell motility assays
Molecular biology: Gene and recombinant DNA cloning, specialty plasmids design, PCR, DNA sequencing analysis, southern blot, DNA isolation, competent cells preparation
Structural biology: Virtual docking and homology modeling, sequences alignment/analysis.
Experience
Feb 20017- Present Alera Labs, LLC Durham, NC
Routine testing and method validation using HPLC, GC, LC-MS and UV-Vis of customer samples (enzymes, polymers, hormones, drugs, vitamins, pigments, etc.) according to approved procedures
Perform stability studies by following the testing calendar, protocols and specified methods to produce accurate results on time
Perform various analytical testing using HPLC, LC-MS, GC, Karl Fischer coulometric titrator, UV-Vis,
Documenting all work as per SOP’s and GLP
Regular calibration and maintenance of analytical equipment (HPLC, LC-MS, UV-Vis, KF, etc)
Apr 2007- June 2015
UNC at Chapel Hill
Chapel Hill, NC
Research Assistant Professor
Demonstrated that GAPDHS, one of the key enzymes of glycolysis in sperm can be a good target for male contraception.
Adapted, validated and performed biochemical enzyme assays and in vivo assays suitable for drug candidates screening using throughput screening format (HTS).
Using pgk2 and gapdhs knockout transgenic mouse models identified significant difference in pattern of tyrosine phosphorylation in sperm of mice.
Performed clinical study in order to identify mutations in sperm of patients suffering from male infertility.
Created database of motility characteristics and metabolic parameters for fertile and infertile male patients of IVF clinics of UNC in purpose of improving male infertility diagnostics and IVF outcome.
Proved the importance of sperm hyperactivation in male infertility diagnostic, the test not currently performed by clinicians.
For the first time solved crystal structures of mouse GAPDHS and human GAPDHS
Jan 2004- Apr 2007
UNC at Chapel Hill
Chapel Hill, NC
Postdoctoral fellow
Expressed and purified mouse and human GAPDHS in large scale.
Using transgenic pgk2 knockout transgenic mouse model determined that PGK2 is required for sperm energy production and motility.
Established enzymes and glycolytic metabolites assays for sperm.
Using metabolomics approaches determined differences in metabolism of sperm lacking PGK2 or GAPDHS.
Demonstrated the impact of single mutation in pgk2 on reproductive performance
June 2002- Jan 2004
UNC at Chapel Hill
Chapel Hill, NC
Postdoctoral fellow
Investigated the role of Sox2 transcriptional factor in nervous system development.
Participated in sox2 knockout mouse model phenotype characterization, mice colony maintenance and breeding studies.
Education
2002
Lomonosov Moscow State University
Moscow, Russia
PhD in Biochemistry
1998
Samara State University
Samara, Russia
Master of Science in Biology
Collaboration with Industry Organizations
GlaxoSmithKline, RTP, NC (2013-2014) Optimized enzymatic bioassay for industry need, supplied company with pure enzymes
Center for Integrative Chemical Biology and Drug Discovery, Chapel Hill, NC (2010-2015)
Performed compound screening in HTS format and analyzed the results
BRITE’s drug discovery center, Durham,NC (2007-2010)
Performed personnel training to do bioassay, setup assay, express enzyme for HTS and troubleshooting procedures for bioassay or enzyme expression
RTI, RTP, NC (2014-2015)
Performed personnel training to do bioassay, set up assay, provided with enzymes for performing HTS and troubleshooting procedures for bioassay
Awards
Lonnie D. Russell Travel Award, first recipient, American Society of Andrology, 2006
Trainee Merit Award, American Society of Andrology, 2006
Young Investigators Award from the Russian Academy of Sciences (UNESCO International Congress), Moscow, Russia, 2000
References
References are available on request.
Publications
Danshina PV, Qu W, Temple B, Rojas RJ, Miley M, Machius M, Betts L, O’Brien DA (2016) Structural analyses to identify selective inhibitors of glyceraldehyde 3-phosphate dehydrogenase-S, a sperm-specific glycolytic enzyme. Mol Hum Reprod 6:410-26.
Lamson DR, House AJ, Danshina PV, Sexton JZ, Sanyang K, O’Brien DA, Yeh LA, Williams KP (2011) Recombinant human sperm-specific glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) is expressed at high yield as an active homotetrameric form in baculovirus-infected insect cells. Protein Expr Purif 75:104-13.
Sexton JZ*, Danshina PV*, Lamson DR, Hughes M, House AJ, Yeh LA, O’Brien DA, Williams KP (2011) Development and implementation of a high throughput screen for the human sperm-specific isoform of glyceraldehyde 3-phosphate dehydrogenase (GAPDHS). Curr Chem Genomics 5:30-41. *joint first authors
Danshina PV, Geyer CB, Dai Q, Goulding EH, Willis WD, Kitto GB, McCarrey JR, Eddy EM, O’Brien DA (2010) Phosphoglycerate kinase 2 (PGK2) is essential for sperm function and male fertility. Biol Reprod 82:136-145.
Danshina PV, Temple B, Williams KP, Sexton JZ, Yeh L, O’Brien DA (2009) New strategies for male fertility control: sperm glycolytic enzymes as targets. J Androl 30 (Suppl):23
Dan'shina PV, Schmalhausen EV, Arutiunov DY, Pleten AP, Muronetz VI (2003) Acceleration of glycolysis in the presence of the non-phosphorylating and the oxidized phosphorylating glyceraldehyde-3-phosphate dehydrogenases. Biochemistry (Mosc) 68(5):593-600
Arutyunova EI, Danshina PV, Domnina LV, Pleten AP, Muronets VI (2003) Oxidation of glyceraldehyde-3-phosphate dehydrogenase enhances its binding to nucleic acids. Biochem Biophys Res Commun 307:547-552
Danshina PV, Schmalhausen EV, Avetisyan AV, Muronets VI (2001) Mildly oxidized glyceraldehyde-3-phosphatase dehydrogenase as a possible regulator of glycolysis. IUBMB Life 51:309-314
Fokina KV, Yazykova MY, Danshina PV, Schmalhausen EV, Muronets VI (2000) Participation of glyceraldehyde-3-phosphate dehydrogenase in the regulation of 2,3-diphosphoglycerate level in erythrocytes. Biochemistry (Mosc) 65:463-468