Ming Dong, BS

ORISE Research Fellow

Laboratory of Molecular Virology

DETTD, CBER/FDA

***** *** ********* ******,

Silver Spring, MD20993-0002

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301-***-****(Cell), 240-***-****(Office)

STATEMENT OF QUALIFICATIONS:

30+ years of research and development experience in immunology, virology, molecular biology, biotechnology and animal study.

In depth knowledge and hands-on experience on immunoassays such as Immunofluorescent antibody technique (IFA); ELISA; ICH and Western blot.

Highly experienced on molecular biology methods including DNA and RNA extraction; DNA cloning; single/multiple mutagenesis; gene expression and protein purification; PCR/RT-PCR and qPCR; DNA sequencing, sequence analysis and Bioinformatics; genotyping by PCR.

Expertise on virology technology including viral infectivity assays, titration, neutralization assay as well as serology, pathology, immunohistochemistry and nucleic acid evaluation for in vitro samples such as cell culture, and in vivo samples from animals.

Expertise on animal studies: handling mouse, rabbit, administrating agents to animal via intradermal, intra-muscular and subcutaneous injection, bleeding mice and rabbits.

Experience in managing research projects and laboratories, coordinating between cooperated labs.

Highly experienced and knowledgeable on cell culture technology including culturing PBMC, protozoan parasites and numerous cell lines.

Experienced on working in the Biohazard level 2 and 3 laboratories and in CDC Select Biological Agents program.

Excellent on experimental design, method development and modification, SOP writing, data recording and interpretation and scientific report writing.

Excellent analytical thinking and judgment skills, and effective trouble shooting skills.

Ability to work independently as well as collaboratively and good at oral and written communication skills and interpersonal skills.

PROFESSIONAL EXPERIENCE AND SKILLS

Oct. 2015 – present ORISE Fellow (Contract biologist), LMV, Division of Emerging and Transfusion Transmitted Diseases, OBRR, CBER, FDA, Silver Spring, MD

Cameroon HIV-1 sample Genotyping and Molecular Epidemiological Investigation

• Extracting HIV-1 RNA from patient plasma; RT-PCR;

• Genotyping HIV-1 Cameroon patient samples using Sanger sequencing and Bioinformatics’ software;

• Testing HIV-1 plasma viral load using Abbott m2000sp and m2000rt;

• Strictly following sterile assurance procedures and GLP regulations in PBMC isolation from whole bloods, cryopreservation and storage;

• Isolating HIV-1 virus from Patient plasma or infected PBMC; Monitor virus replication by HIV-1 p24 ELISA ;

Sept. 2013 – Sept. 2015, ORISE Fellow (Contract biologist), LEP, Division of Emerging and Transfusion Transmitted Diseases, CBER/FDA, Silver Spring, MD Standardizing methods for spiking low prevalence pathogens into Healthy donor and patient blood and plasma to assist diagnostics developers with performing clinical sensitivity studies required for FDA clearance of diagnostic devices.

• Established culture conditions, identified appropriate cryopreservation method for sample pathogen (Bacillus anthracis; Francisis tularensis; Yersinia pseudotuberculosis; Dengue virus; Leishmania donovani; Babiesia microti; Escherichia coli and Staphylococcus aureus including 4 NIAID Category A pathogens)

• Developed singleplex qPCR., RT-qPCR for each pathogen;

• Developed and updated Standard Operating Procedures (SOPs) for procedures and work instructions of the methods and equipment’s calibration, maintenance and operations.

• Performed quality control testing of in house validation by qPCR., RT-qPCR strictly followed SOP and sent spiking sample DNAs and RNAs to contractor for external validation to test the stability and re-productivity of this standardized method;

• Performed qualification, preventive maintenance and calibration of the equipment used in the testing such as real time PCR instrument, Automated cell counter, microscope etc.

• Managed and performed qualification of all types of reagents used.

• Recorded experimental observation, interpreted experimental data and wrote technical reports.

• Contributed to handling lab inventory and management and developing, evaluating and standardization of protocols for research purposes.

Dec. 2003-Aug. 2013, Research Associate II, Henry M. Jackson Foundation/USUHS, Department of Preventive medicine and Biometrics,

HIV-1 vaccine study in mouse, rabbit and rhesus monkey delivered by VEE system or HIV-1 gp140 protein with multiple regimes and adjutants.

• Prepared HIV-1 vaccine for animal study. HIV-1 Clade B, C, D, E envelop gene-gp160, gp140, gp160 CT were cloned into Venezuelan Equine Encephalitis Virus (VEE) replicon expression system. VEE replicons with HIV-1 gene were in vitro transcribed and transfected cell by electroporation, harvested by ultracentrifuge.

• Conducted and coordinated animal study.

• Tested sera-conversion of the animal study by Three-step sandwich ELISA.

• Tested cross-reactive neutralization antibody using HIV-1 pseudotyped viruses from panel of clade A, B, C and some primary isolated clade E and D;

• Studied the character of broadly neutralizing antibodies from vaccinated animals (rabbit and monkey) by competing Neutralization Assay using CD4bs glycoprotein, glycan-specific neutralizing antibody by NA with modified HIV-1 pseudotyped viruses (added and eliminated glycosylation site on envelope protein.)

• Developed an assay for measuring HIV-1 neutralizing antibody using SIV mutant by introducing the HIV-1 membrane proximal neutralization epitopes screen human blood samples to find broad neutralization antibody.

• Operated, calibrated and maintained Luminometer, Fluorescent microscope and other equitment

• Planned and developed research studies by choosing the methods and procedures, searched literature. Conducted experimental studies, analyzed data, and summarized findings for papers and reports.

• Help managing the lab by ordering supply and calibrating, maintaining equitment. Contributed to

Dec.1999-Dec.2003 Research Associate I, Henry M. Jackson Foundation/USUHS, Department of Preventive medicine and Biometrics,

Induction of Primary Virus-Cross-Reactive Human Immunodeficiency Virus Type 1-Neutralizing Antibodies in Small Animals by Using an Alphavirus-Derived In Vitro Expression System

• Conducted and coordinated animal study.

• Tested sera-conversion of the animal study by ELISA.

• Test cross-reactive neutralization antibody using HIV-1 pseudotyped viruses.

• Cloned HIV-1 envelop gene- gp160, gp140, gp160 CT into Venezuelan Equine Encephalitis Virus (VEE) replicon expression system-cloning, screening, site-directed mutagenisis, sequencing and western blot for gene expression.

June 1998-Dec.1999 Research assistant, Henry M. Jackson Foundation/USUHS, Department of Preventive medicine and Biometrics,

Induction of Primary Virus-Cross-Reactive Human Immunodeficiency Virus Type 1-Neutralizing Antibodies in Small Animals by Using an Alphavirus-Derived In Vitro Expression System

• Cloned HIV-1 envelop gene into VEE replicons and characterized with RT-PCR technique, West blot and ICH

• Conducted experiments by using Venezuelan Equine Encephalitis Virus (VEE) replicons expression system study neutralization domains on the HIV-1 envelopes and neutralization antibodies.

Sept. 1996-June 1998 Research fellow, Division of Virology, National Vaccine and Serum Institute, Beijing, China

• Purification of HIV-1 gp120 proteins, which was expressed in the Baculovirus Expression Vector System (BEVS).

• Chicken embryo live Yellow Fever Virus vaccine production.

Sept. 1995-Sept. 1996 Visiting Scientist, Department of Preventive medicine and Biometrics, Uniformed Services University of the health Sciences

Cloned and characterized the envelope genes from different patient's lymphocyte with RT-PCR technique.

March 1995-Sept.1995 WHO FELLOW, Department of Preventive medicine and Biometrics, Uniformed Services University of the health Sciences

• Focused on the expression and characterization of HIV-1 envelopes protein associated with a broadly reactive neutralizing antibody response.

• Compared different transfection techniques to achieve higher titer of HIV-1 pseudotype viruses for Neutralization Assay.

Sept. 1985-March 1993, Research Assistant, Division of Virology, National Vaccine and Serum Institute, Beijing, China

Using molecular Biology techniques to develop vaccine and bioproduct.

• Worked on the expression of synthesized Human Granulocyte Colony Stimulating Factor (hG-CSF) in the Baculovirus Expression Vector System (BEVS).

• Cloned and expression of HIV-1 gp120, gp160, gp41 and p24 in the BEVS.

• Conducted experiments of DNA synthesis, cloning, sequencing, site-directed mutagenesis and expression of human G-CSF gene.

• Cloned and expression of Hepatitis C Virus E1, E2 gene by RT-PCR.

• Constructed high-level expression vector PKBS.

• Involved in the study of Herpes Simplex Virus type I subunit vaccine and Varicella-zoster Virus attenuated vaccine.

TECHNICAL SKILLS

Immunology and Virology: In depth knowledge and hands-on experience on immunoassays such IFA; ELISA; ICH and Western blot. Expertise on virology technology including viral infectivity assays, titration, neutralization assay as well as serology, pathology, immunohistochemistry and nucleic acid evaluation for in vitro samples such as cell culture, and in vivo samples from animals.

Vaccine research: Experience in studying B cell responses on mice, rabbit and rhesus monkeys immunized with HIV vaccine

Infectious diseases: Experience in handling human and animal pathogens like HIV/SIV, VEE, HSV-1, HBV and VZV and infectious disease diagnosis. Extensive experience in handling of blood and blood products from mice, monkeys and humans and handling all clinical samples for testing. Experienced on working in the Biohazard level 2 and 3 laboratories and in CDC Select Biological Agents program.

Molecular Biology: Genomic and plasmid DNA isolation, RNA isolation, DNA sequencing, Real-time PCR and gel electrophoresis. Primer designing, PCR, cloning, development prokaryotic and eukaryotic expression constructs. single/multiple site –directed mutagenesis. Knowledge of DNA/RNA sequence analysis, multiple sequence alignment and use of bioinformatics techniques and software. RT-PCR and SDS/PAGE and western blots and protein purification

Cell Culture and Tissue culture: Highly experienced and knowledgeable on cell culture technology including culturing PBMC, protozoan parasites and numerous cell lines which includes both infectious and non-infectious cell lines. Knowledge of maintaining suspension and adherent cell lines, storage and handling

Animal Experimentation: handling mouse, rabbit, administrating agents to animal via intradermal, intra-muscular and subcutaneous injection, bleeding mice and rabbits.

Lab management: Knowledge of handling lab inventory and management and developing, evaluating and standardization of protocols for research purposes with over 10 years’ experience. Applying technical expertise to conduct scientific review and evaluation of data, test methods and procedures, and reference methods. Ordering reagents, equipment and new instrumentation for entire lab. Negotiated with vendors to ensure cost savings. Organized reagents and equipment. Contributed to management of the lab budget. Involving coordinating between cooperated labs.

EDUCATION AND TRAINING

Bachelor's Degree in Marine Biology, 1981-1985, Ocean University of China, Qingdao, China.

CBER New Reviewer Training, June 2015, FDA, Silver Spring, MD.

“Molecular Biology”, “A Course in Laboratory Diagnosis of Viral Infection”, “Application of Molecular Biological Techniques”, and “Molecular Medical Genetics courses”. Between 1986 and 1990, The Sino-Danish Postgraduate Biomedical Training Center, Beijing, China

“The Large Scale Mammalian Tissue Culture course”. 1990, The Eastern China Chemical Engineering Institute, Shanghai, China

“Advanced English Language Training Course for WHO Fellowship”, 1993, The English Training Center, Hunan Medical University, Changsha, Hunan Province, China,

AWARDS

World Health Organization Fellowship Award, 1995

Department of Preventive Medicine and Biometrics, Edward F Herbert School of Medicine, Uniformed Services University of the Health Sciences Bethesda, MD

PUBLICATIONS

M. Dong, C. Fisher, G. Anez, M. Rios, H.L. Nakhasi, J.P. Hobson, M. Beanan, D. Hockman, E. Grigorenko, and R. Duncan. 2016. Standardized methods to generate mock (spiked) clinical specimens by spiking blood or plasma with cultured pathogens. J Appl Microbiol. 120:1119-1129.

Duncan, R., M. Kourout, E. Grigorenko, C. Fisher, and M. Dong. 2016. Advances in multiplex nucleic acid diagnostics for blood-borne pathogens: promises and pitfalls. Expert Rev Mol Diagn. 16:83-95.

Hockman, D., M. Dong, H. Zheng, S. Kumar, M.D. Huff, E. Grigorenko, M. Beanan, and R. Duncan. 2016. Comparison of multiplex PCR hybridization-based and singleplex real-time PCR-based assays for detection of low prevalence pathogens in spiked samples. J Microbiol Methods. 132:76-82.

Quinnan, G.V., Jr., O. Onabajo, P. Zhang, L. Yan, J.J. Mattapallil, Z. Zhang, M. Dong, M. Lu, D. Montefiori, C. LaBranche, and C.C. Broder. 2014. Immunization of rabbits with highly purified, soluble, trimeric human immunodeficiency virus type 1 envelope glycoprotein induces a vigorous B cell response and broadly cross-reactive neutralization. PLoS One. 9:e98060.

Quinnan, G.V., Jr., P. Zhang, M. Dong, H. Chen, Y.R. Feng, M. Lewis, and C.C. Broder. 2013. Neutralizing antibody responses in macaques induced by human immunodeficiency virus type 1 monovalent or trivalent envelope glycoproteins. PLoS One. 8:e59803.

Zhang, P.F., F. Cham, M. Dong, A. Choudhary, P. Bouma, Z. Zhang, Y. Shao, Y.R. Feng, L. Wang, N. Mathy, G. Voss, C.C. Broder, and G.V. Quinnan, Jr. 2007. Extensively cross-reactive anti-HIV-1 neutralizing antibodies induced by gp140 immunization. Proc Natl Acad Sci U S A. 104:101**-*****.

Quinnan, G.V., Jr., X.F. Yu, M.G. Lewis, P.F. Zhang, G. Sutter, P. Silvera, M. Dong, A. Choudhary, P.T. Sarkis, P. Bouma, Z. Zhang, D.C. Montefiori, T.C. Vancott, and C.C. Broder. 2005. Protection of rhesus monkeys against infection with minimally pathogenic simian-human immunodeficiency virus: correlations with neutralizing antibodies and cytotoxic T cells. J Virol. 79:3358-3369.

M. Dong, P.F. Zhang, F. Grieder, J. Lee, G. Krishnamurthy, T. VanCott, C. Broder, V.R. Polonis, X.F. Yu, Y. Shao, D. Faix, P. Valente, and G.V. Quinnan, Jr. 2003. Induction of primary virus-cross-reactive human immunodeficiency virus type 1-neutralizing antibodies in small animals by using an alphavirus-derived in vivo expression system. J Virol. 77:3119-3130.

Quinnan, G.V., Jr., P.F. Zhang, D.W. Fu, M. Dong, and H.J. Alter. 1999. Expression and characterization of HIV type 1 envelope protein associated with a broadly reactive neutralizing antibody response. AIDS Res Hum Retroviruses. 15:561-570.

Quinnan, G.V., Jr., P.F. Zhang, D.W. Fu, M. Dong, and J.B. Margolick. 1998. Evolution of neutralizing antibody response against HIV type 1 virions and pseudovirions in multicenter AIDS cohort study participants. AIDS Res Hum Retroviruses. 14:939-949.

M. Dong, Y. Xie, J. Xiang et al .1997. Expression of HIV-1 Env protein in Insect Cells and Evaluation of Their Immunological Properties. Chin J Microbiol Immunol, Vol. 17, No. 1

Y. Xie, M. Dong, P.F. Zhang. 1997. Expression of HIV-1 p24 protein in Insect Cells and Evaluation of Their Immunological Properties. Chines J of Virology, Vol. 13, No.3

H. Liu, C. Guo, W. Wei, M. Dong. 1997. Studies on the Molecular Epidemiology of Poliovirus Type 1 in China. Chin J Epidemiol. Vol. 18, No. 4

C. Pang, X. Liu, P. Peng, M. Dong, G. Wang and Y. Wang. 1995. Application of Assaying VZV Ig G by ELISA. Chinese Journal of Biological. 8(1):23

M. Dong and G. Jing, 1990. A Simple and Efficient Method for Isolation DNA with Molecular Weight and High Purity from Cultured Cells Progress in Biochemistry and Biophysics Vol. 3.

Y. Zhang, M. Dong, X. Sun, Z. Tong and Y.Wang, 1989. Study of Herpes Simplex Virus Type I (HSV-1) Subunit Vaccine. I. Extraction of Subunit Polypeptide. Chinese Journal of Microbiology and Immunology Vol. 9, No.1

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REFERENCES

Indira K Hewlett, Ph.D.

Chief, Laboratory of Molecular Virology

Division of Emerging and Transfusion Transmitted Diseases, CBER/FDA

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

240-***-****

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Robert C. Duncan, Ph.D.

Division of Emerging and Transfusion Transmitted Diseases, CBER/FDA

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

240-***-****

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Gerald V. Quinnan, Jr., MD, RADM (Ret), USPHS

Professor Emeritus

F. Edward Hébert School of Medicine

Uniformed Services University of the Health Sciences

4301 Jones Bridge Road

Bethesda, MD 20814

301-***-**** [Cell]

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