Fushan Wang, Ph.D., Johnson and Johnson company

Tel: 610-***-****(Cell); Email: adutvh@r.postjobfree.com; adutvh@r.postjobfree.com

Profile

Highly motivated senior scientist with about ten years’ experience in both upstream production and downstream purification of various gene therapy vectors (lentivirus/adenovirus/adeno-associated virus/oncolytic HSV-1), viruses for viral clearance study from bench to production scales, and protein(HSV glycoproteins) and antibody(anti-HSV glycoprotein)production and purification under GLP/GMP

Expertise involved into the above processes ranges from development of upstream high yield serum-free growth methods (Hyperflask, CellStack, Roller bottle and bioreactor) to downstream purifications for AAV, lentivirus, X-MuLV, mice minute virus (MVM), pseudorabies virus (PrV), herpes simplex virus-1 (HSV-1), and Reo-3 virus, including liquid chromatography (affinity, HIC, IEX, and SEC) on AKTA FPLC systems (Prime, Purifier, Explorer, and Avant), depth filtration, tangential flow filtration (TFF) and diafiltration (Repligen Krosflo KR2i system)

Strong background in virology demonstrated in making various recombinant viruses (baculovirus, adenovirus, adeno-associated virus, retrovirus, and herpes simplex virus)

Experienced in HSV-2 vaccine development for about 10 years

Demonstrated command of basic and most of the main advanced Molecular Biology/Protein Chemistry techniques spanning from particle size and polydisperity analysis by dynamic light scattering (DLS), quantitative real time PCR, Western blotting, baculoviral protein expression system, to a variety of immunological assays (ELISA, ELISPOT and FACS) and virological methods

Strong qualifications also include interpreting research data, preparing research reports for presentation and publications, finalizing SOPs and batch records, well-developed supervisory, interpersonal and communication skills, and excellent problem-solving and decision-making capability.

The final recently purified oncolytic HSV in JNJ meets FDA guidelines in host cellular DNA and protein limits qualified for clinical trial.

Professional Experience

Johnson and Johnson in Rockville, MD (July 2018 –Present)

Senior Scientist

Led and implemented the development of both upstream production and downstream purification of oncolytic HSV-1, and routinely performed the upstream production and downstream purification of viruses using Capto Core 700 chromatography under GMP

1)Development of upstream production of oHSV-1 in serum-free medium using Hyperflask, CellStack and Bioreactor

2)Improvement of Mustang Q anion exchange downstream purification to increase final viral yield from 6% to 63%;

3)Development of Capto Core 700 chromatography from initial to scale up runs (overall yield 86%), including depth filtration, tangential flow filtration (TFF) and diafiltration (Repligen Krosflo KR2i system); carrying out a variety of virological and molecular analytical methods to qualify the final purification process;

4)Routine weekly implementation of production and purification of viruses under GMP

University of Pennsylvania (March 2017 – June 2018)

Senior Research Investigator – Infectious Disease Division

HSV-2 subunit vaccine development – production in bioreactor by baculoviral expression system and purification of glycoproteins gC, gD and gE of HSV-2 and antibodies anti-gC, gD and gE by chromatography; efficacy and safety assessments by quantitative real time PCR and ELISA

WuXi AppTec - Philadelphia (Feb. 2011 – Jan. 2017)

Senior Scientist - Purification and process development

Served as lead for process development designing upstream growth conditions and novel purification processes for viruses, as well as optimizing existing purification strategies

Process development, production, and purification of gene therapy vectors

1)VSV-G pseudotyped lentivirus: co-transfection of four mix by PEI in 293T cells, infectious titer determination and genomic copy number quantification by real-time PCR, and purification by anion exchange/gel filtration chromatography

2)Adeno-associated virus (AAV): two rHSV-based AAV production and purification by anion-exchange/gel filtration

Production and purification of viruses for viral clearance study

1)Development of high yield serum-free growth methods for X-MuLV, mice minute virus (MVM), pseudorabies virus (PrV), herpes simplex virus-1(HSV-1), and Reo-3 virus in Hyperflask, CellStack, Roller bottle and bioreactor

2)Development of down-stream chromatography purification of best-in-industry viral stocks

X-MuLV, first ceramic hydroxyapatite(CHT) capture column/second gel filtration impurity removal column/third CHT volume reduction column; PrV and HSV-1, first ceramic hydroxyapatite(CHT) capture column/second Capto Core 700 impurity removal column; and Reo-3, first DEAE anion exchange capture column/ second Capto Core 700 impurity removal column

3)Establishment of dynamic light scattering (DLS) for analysis of virus purity and mono-dispersity and routine client DLS analysis services and qualification of viral stocks for viral clearance by solvent detergent, low pH hold inactivation, and nanofiltration

University of Pennsylvania (Aug. 2007–Jan. 2011)

Senior Research Investigator – Infectious Disease Division

Production of glycoproteins by baculovirus expression system and purification of HSV-1/2 subunit vaccine by chromatography; development of HSV-1 and HSV-2 attenuated live virus vaccine and subunit vaccine using mouse and guinea pig models: constructed and chromatography purified HSV-1/-2 glycoprotein E deletion live attenuated virus vaccines; evaluated immunogenicity and efficacy of the vaccines using a variety of immunological and virological methods; and obtained a co-inventor of a patent “Methods of Use for HSV-1 and HSV-2 Vaccines (US20090246227)”

Thomas Jefferson University (June 2006 – Jul. 2007)

Research Assistant – Infectious Diseases Division

Developed an ultrasensitive immunoassay of botulinum neurotoxin by immune-real time PCR and study of pharmacokinetics of botulinum neurotoxin in mice

University of Pennsylvania (Feb. 2002 – May 2006)

Research Specialist – Infectious Diseases Division

Studied gE deletion recombinant HSV-1 neuronal transport using compartmented culture of primary SCG neurons, ophthalmic microinjection techniques and retina and brain immunohistochemistry

University of Pennsylvania (Jan. 2000 – 2002)

Postdoc Fellow-Department of Physiology (connexin intracellular trafficking)

Education

Ph.D., Molecular Biology and Virology, Department of Virology, Wuhan University, China

Publications

1. Wang F., E.E. Brittle, J. Huang, H. Si, and H. M. Friedman. 2010. Herpes simplex virus type 2 glycoprotein E is required for efficient virus spread from epithelial cells to neurons and for targeting viral proteins from the neuron cell body into axons. Virology. 45(2): 269-279.

2. Wang, F., W. Tang, H. M. McGraw, J. Bennett, L. W. Enquist, and H. M. Friedman. 2005. Herpes simplex virus type 1 glycoprotein E is required for axonal localization of capsid, tegument, and membrane glycoproteins. J. Virol.79:13362-72.

3. Wang, F., B. Daugherty, L. Keise, Z. Wei, J.P. Foley, R. C. Savani and M. Koval. 2003. Heterogeneity of claudin expression by alveolar epithelial cells. American Journal of Respiratory Cell & Molecular Biology 29(1):62-7.

4. Brittle, E. E., F. Wang, J. M. Lubinski, R. M. Bunte, and H. M. Friedman. 2008. A replication-competent, neuronal spread-defective, live attenuated herpes simplex virus type 1 vaccine. J. Virol., 82:8431-41.

5. Sama J., F. Wang, and M. Koval 2002. Targeted gap junction protein constructs reveal connexin-specific differences in oligomerization. Journal of Biochemistry 277(23):20911-8.

6. Sarma J., R. A. Meyer, F. Wang, V. Abraham, C. W. Lo and M. Koval. 2001. Multimeric connexin interactions prior to the trans-Golgi network. Journal of Cell Science 114: 4013-4024.

7. Awasthi S, Hook LM, Pardi N, Wang F, Myles A, Cancro MP, Cohen GH, Weissman D, Friedman HM. 2019. Nucleoside-modified mRNA encoding HSV-2 glycoproteins C, D, and E prevents clinical and subclinical genital herpes. Sci Immunol. 2019 Sep 20;4(39): eaaw7083. doi: 10.1126/sciimmunol. aaw7083.

8. Awasthi S, Zumbrun EE, Si H, Wang F, Shaw CE, Cai M, Lubinski JM, Barrett SM, Balliet JW, Flynn JA, Casimiro DR, Bryan JT, and Friedman HM. 2012. Live attenuated herpes simplex virus 2 glycoprotein E deletion mutant as a vaccine candidate defective in neuronal spread. J Virol. 86(8):4586-98.

9. Awasthi S, Lubinski JM, Shaw CE, Barrett SM, Cai M, Wang F, Betts M, Kingsley S, Distefano DJ, Balliet JW, Flynn JA, Casimiro DR, Bryan JT, Friedman HM. 2011. Immunization with a vaccine combining herpes simplex virus 2 (HSV-2) glycoprotein C (gC) and gD subunits improves the protection of dorsal root ganglia in mice and reduces the frequency of recurrent vaginal shedding of HSV-2 DNA in guinea pigs compared to immunization with gD alone. J Virol. 85(20):10472-86.

10. Lubinski JM, Lazear HM, Awasthi S, Wang F, Friedman HM. 2011. The herpes simplex virus 1 IgG fc receptor blocks antibody-mediated complement activation and antibody-dependent cellular cytotoxicity in vivo. J Virol. 85(7):3239-49.

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