Flow Cytometry Clinical Research

Curriculum Vitae

SUJATA PRASAD, Ph.D.

Scientist

Clinical,Translational, & Pre-clinical

MLM Medical Labs, Oakdale, Minnesota

Email: ********@*****.***

Mobile # 612-***-****

Area of specialization: Immunology and Microbiology

Professional Summary

Experienced Cell Culture and Flow Cytometry Scientist with over 10 years in the life sciences industry, specializing in assay development, immune profiling, and advanced cytometry techniques for translational research. Demonstrated expertise in primary, immortalized cell culture and mammalian cell culture such as HEK293, including the expansion and differentiation of cell lines and immune cells under aseptic conditions. Skilled in the design and optimization of multicolor flow cytometry panels for phenotypic and functional analysis of complex cell populations, with proven success in validating panels for applications such as immune checkpoint inhibition, CAR-T cell therapy, and phospho-flow studies.

Adept at data interpretation and statistical analysis, translating findings into actionable insights for preclinical and clinical studies. Known for maintaining high standards in laboratory protocols, troubleshooting technical issues, and ensuring accuracy and reproducibility in data reporting. Strong collaborator with experience in cross-functional teams, regulatory documentation, and client communication, committed to driving innovation and delivering quality results that support critical research objectives and therapeutic advancements. knowledge of regulatory requirements (ICH-GCP) and adept in using cutting-edge technologies to drive innovation in clinical trials.

My proficiency in data management, statistical analysis, and scientific writing allows me to effectively communicate research findings through comprehensive reports, publications, and presentations. I am adept at collaborating with investigators, sponsors and industry partners to facilitate seamless study execution and foster meaningful collaborations.

Core Skills

Optimizing cell Mammalian Cell Culture conditions, and Stable Cell Line Development

Process Optimization and Scale-Up

Regulatory Compliance (GxP, GLP)

performed routine passaging, seeding, and cryopreservation of primary cell lines such as HEK293 and immune cells such as PBMCs, T cells and Treg cells while maintaining sterility in a BSL-2 environment.

Experience with wide variety of immune cells such as T cells, B cells, NK cells, dendritic cells, other myeloid cells, tissue resident memory cells and Treg cells

Ex-vivo cell stimulation using specific stimuli like antigens, cytokines or other activating agents, cell proliferation assays, up-regulation of activation markers, effector cytokine production and cytotoxicity.

Team Collaboration and Technical Training

Experience in designing and validation of protocols in strict compliance with GCP and GCLP guidelines.

Provide scientific expertise in the design of protocols and presentation of scientific data to investigators and sponsors.

Strong scientific skills and ability to effectively communicate these aspects in writing (Abstract and manuscript preparation).

Applied knowledge on clinical research, study design, statistical analysis, and medical terminologies.

Experience in scientific publication, authoring and reviewing clinical study protocols, analyzing, evaluating clinical, pre-clinical and translation data gathered during the study and preparing study reports.

Provided scientific input to the pre-clinical and translational team for protocol development and technical supervision.

Offered editorial and peer reviewed services for several clinical research articles.

Proven track record in the field of immunology, scientific writing, present research outcomes in the form of journal manuscripts, grants, reports, posters, and oral presentation.

Strong understanding of immune system functions and dysregulation under various disease conditions, solid understanding of immunotherapeutic areas and quickly develop expertise in new therapeutic areas.

Perform literature reviews to assist with data interpretation and preparing results.

Excellent computer skills to include MS Word, MS Excel, MS PowerPoint, MS outlook, Teams and Adobe Acrobat, GraphPad.

Understanding of JReview to analyze clinical data.

Understanding of the drug development process in all phases of testing and adverse events.

Demonstrate creative and novel approaches to solving scientific problems.

Proven administration and organizational skills, multitasking, project management, teamwork, and ability to quickly acquire knowledge in a fast pace working environment.

Demonstrate the ability to meet project goals as a study director/lead scientist.

My skill set encompasses a wide range of scientific activities. I am proficient in scientific writing, having successfully authored and published more than 23 research manuscripts in esteemed peerreviewed journals. These publications have garnered significant recognition, with 385 citations to date. Additionally, I have shared my research findings through 23 oral presentations and abstract submissions, effectively disseminating knowledge within the scientific community.

Relevant Work Experience

March 2022 – Till date

Scientist, MLM Medical Labs. I focus on scientific writing, authoring, and reviewing pre-clinical, translational, and clinical study protocols, designing and executing experimental studies, analyzing, evaluating data gathered during the study and preparing study reports. I provide scientific expertise in the development of new business pitches and proposals. In addition to providing scientific inputs, I perform routine systematic literature reviews for designing and validation of protocols in strict compliance with GCP and GCLP guidelines. I am involved in conducting research on diverse pre-clinical disease models, including imiquimod (IMQ)induced Psoriasis, lung fibrosis, and inflammatory bowel disease. In my role as a study director, I oversee numerous clinical studies and actively contribute to the designing of flow cytometry and cell culture based-based investigations. I performed routine passaging, seeding, and cryopreservation of primary cell lines such as HEK293 and immune cells such as PBMCs, T cells and Treg cells while maintaining sterility in a BSL-2 environment. My work revolves around assessing the effects of specific test article treatments in these disease models, aiming to uncover potential therapeutic interventions.

I possess extensive expertise in the field of immunomodulation. With a deep understanding of the complex interactions between the immune system and various diseases or conditions, I am well-versed in the mechanisms and strategies employed to modulate immune responses. As part of my responsibilities, I collaborate on designing flow cytometry-based investigations.

I am skilled in the design and implementation of immunomodulatory therapies, such as immune checkpoint inhibitors, cytokine therapies, vaccination strategies, acetylation, phosphorylation assays and targeted immunotherapies.

My proficiency extends to critically analyzing immunological data, interpreting results, and drawing meaningful conclusions.

Technical skills: More than 10 years of experience in Flow Cytometry, Flow panel designing, cell culture-maintaining cell lines, ensuring optimal growth conditions for accurate experimental outcomes, M1/M2 phenotyping using THP-1 cell Line. Ex-vivo assays in response to sponsors test articles, Ex-vivo cell stimulation using specific stimuli like antigens, cytokines or other activating agents, cell proliferation assays, upregulation of activation markers, effector cytokine production, cytotoxicity, MTT assays, alamar blue assays, ELISA data analysis, qPCR data analysis, Protocol Validation, animal handling and clinical scoring.

Software skills: FlowJo, GraphPad Prism, MS Word, MS Excel, MS PowerPoint, MS outlook, and Adobe Acrobat to analyze and visualize scientific data accurately. Trained in high-dimensional data analysis techniques, including t-SNE, vi-SNE, phenograph, Cytobank, and FlowSOM using FlowJo Plugin programs. This expertise allows for comprehensive exploration and interpretation of complex datasets.

April 2021 – March 2022

Research Scientist, Department of Medicine, Division of Infectious Disease and International Medicine, University of Minnesota, MN. In this role I investigated neuroimmune responses to viral infection and development of immunological protection to re-infection, specifically investigated the development of Tissue resident memory T cells within the infected brain and its role in combating infection employing Flow cytometry as a key tool in my research.

Moreover, my publications showcase the meticulous experimental design and rigorous data analysis methodologies employed in my research. I have utilized a range of techniques, including animal based research, in vitro and ex vivo studies, Immunemodulation, molecular biology assays, and immunohistochemistry, to investigate the impact of neuroimmunomodulation in diverse contexts.

Through my scientific publications and presentation, I have effectively communicated my findings of neuroimmunomodulation to the scientific community.

Technical skills: Flow Cytometry (more than 8 Colors), microglial cell culture, cell isolation from brain, spleen, lymph nodes. Sorting and adoptive transfer of specific immune cells. Immune response generated by T cells, Regulatory T cells and Tissue resident memory T cells (TRM cells), T cell activation, expansion, survival, cytokine production, and cell cytotoxicity. Ex-vivo cell stimulation using specific stimuli like specific peptides. Experiments involving immune checkpoint inhibitors such blocking of PD1: PD-L1 pathway. Neuroimmune modulation by T cell therapy.

Software skills: FlowJo, GraphPad Prism, MS Word, MS Excel, MS PowerPoint, MS outlook, and

Adobe Acrobat, trained in high dimensional data analysis- t-SNE, vi-SNE, phenograph, Cytobank and FlowSOM using FlowJo Plugin programs. I am well trained in high dimensional data analysis utilizing programs such as t-SNE, vi-SNE, phenograph, Cytobank, and FlowSOM, which are valuable plugins within the FlowJo software.

October 2014 14 April 2021

Postdoctoral associate, Department of Medicine, Division of Infectious Disease and International Medicine, University of Minnesota, MN. In this role, I dedicated my efforts to comprehending the host defense mechanisms against cytomegalovirus (CMV) infections in the brain. A particular focus of my investigation has been on studying the infiltration of peripheral lymphocytes into the brain and their interactions with glial cells.

Throughout my work, I have employed a variety of techniques and methodologies. Flow cytometry has been an essential tool, allowing me to perform flow-based analysis of immune subsets present in diverse samples, including brain tissue, peripheral blood, cell lines, bronchial alveolar lavage, and pleural fluid. This analytical approach has enabled me to investigate the activation, proliferation, and cytokine production of these immune subsets accurately.

In addition to flow cytometry, I have utilized cell culture techniques to study specific cell types. This includes the culture and isolation of microglial cells, as well as mononuclear cells from various sources. Furthermore, I have employed chemotaxis assays and cell sorting methodologies to investigate cellular migration and isolate specific immune populations.

Technical skills: Handling small animals, intracerebroventricular injection, tail vein injection, intraperitoneal injection, and perfusion. Flow Cytometry and Cell Culture: Microglial cell culture, isolation of mononuclear cells from Brain, peripheral blood, sputum, bronchial alveolar lavage and pleural fluid, chemotaxis assay and cell sorting. Flow based analysis of T, B and other immune cell subsets, intracellular cytokines, Immunohistochemistry. Molecular Biology: Plasmid isolation, cloning, DNA from Blood & Urine, PCR/RT-PCR, RNA Purification and c-DNA Synthesis, SSCP, Agarose gel Electrophoresis PAGE (native & SDS), Western blotting.

Software skills: FlowJo, GraphPad Prism, MS Word, MS Excel, MS PowerPoint, and Adobe Acrobat

Certifications

Introduction to Systematic Review and Meta Analysis

Clinical Trials Management and Advanced Operations

Good Clinical Practice Guidelines/ICH

Good Clinical Laboratory Practice

Select Agents and Toxins Regulation

Animal Biosafety and Risk Assessment

AALAS Certification

Education

Ph.D. 2013, in Microbiology, All India Institute of Medical Sciences, New Delhi, India

Thesis Title: Characterization and clinical implications of Staphylococcus epidermidis biofilms in device related infections

MSc. 2003, Vikram University, Ujjain, M.P. India

BSc. 2000, Rani Durgawati University, Jabalpur, M.P. India

Professional Membership

1.American society of Virology 2019-2020

2.International Society for Neurovirology 2015-2019

3.The Association for Research in Vision and Ophthalmology 2012-2013.

4.American Society of Microbiology 2012-2013

Editor

1.Editorial board member of Acta Scientific Neurology

2.Academic Editor of International Journal of Microbiology, Hindawi Limited 3rd Floor, Adam House 1 Fitzroy Square, London W1T 5HF. I have edited the following manuscript:

a.International Journal of Microbiology, Manuscript ID- ID 8840561, Title “Prevalence of E. coli ST131 among Uropathogenic E. coli Isolates from Iraqi Patients in Wasit Province/Iraq”.

b.International Journal of Microbiology, Manuscript ID- ID 8845835, Title “Assessment of

Pertussis Vaccine Protective Effectiveness in Children in the Amhara Regional State, Ethiopia”.

c.International Journal of Microbiology, Manuscript ID- ID 8845835, Title “The Role of T helper 17 (Th17) and regulatory T cells (Treg) in the Pathogenesis of Vulvo-Vaginal Candidiasis among HIV-infected Women”.

d.International Journal of Microbiology, Manuscript ID- ID 8889086, Title “Evaluation of Procalcitonin, C-reactive protein and Interleukin-6 as Early Markers for Diagnosis of Neonatal Sepsis”.

Reviewer (Total 7)

1.Frontiers in Cellular and Infection Microbiology, Review Report – 823262, Title “ Gut microbiome alterations in patients with visceral obesity based on QCT.

2.Immunity, Inflammation and Disease Manuscript ID IID3-2020-09-0264, Titled "Does COVID-19 contribute to development of neurological disease?"

3.Journal: Brain Behavior and Immunity Ref: BBI_2020_1803_R1

Title: DHA Attenuated Japanese Encephalitis Virus Infection-Induced Neuroinflammation and Neuronal Cell Death in Cultured Rat Neuron/glia

4.Reviewed manuscript for volume 3 titled “Relationship between Probiotics and Gut-Skin Axis in Skin Wound Healing: a Recent Update” in multivolume book entitled “Probiotic Research in Therapeutics” with Springer Nature.

5.Scientific Reports Ref 82e4eb39-d400-457d-b525-22ae9e2e1d55: Title:

SPARC coordinates extracellular matrix remodeling and efficient recruitment to and migration of antigen specific T cells in the brain following infection.

6.Microbial Pathogenesis Ref: YMPAT_2018_1210

Title: Frequencies of PD-1- and PD-L1- positive T CD3+CD4+, T CD3+CD8+ and B CD19+ lymphocytes and its correlations with other immune cells in patients with recurrent furunculosis.

Travel Awards

1.Travel award from Department of Science and technology (DST), INDIA to attend The Association of Research in Vision and Ophthalmology (ARVO)-2012 annual meeting.

2.Travel Bursary from IMMUNOLOGY FOUNDATION to attend ARVO-2012

Research Publications and Editorial- (Total no: 24) (Total citation: 385)

1.Shilpi Singh, Debashis Barik, Karl Lawrie, Iteeshree Mohapatra, Sujata Prasad, Afsar R Naqvi, Amar Singh, Gatikrushna Singh (International Journal of Medical Sciences 2023) Unveiling Novel

Avenues in mTOR-Targeted Therapeutics: Advancements in Glioblastoma Treatment

(https://doi.org/10.3390/ijms241914960)

2.Shilpi Singh, Debashis Barik, Ananta Prasad Arukha, Sujata Prasad, Iteeshree Mohapatra, Amar Singh, Gatikrushna Singh (Biomedicines 2023) Small molecules targeting new immune soldier: A novel approach forcancer treatment (https://doi.org/10.3390/biomedicines11102621).

3.Amar Singh, Sujata Prasad, Gatikrushna Singh (2023) Microbiome mediated T cell regulation, Inflammation and disease. Book Chapter, Microbiome Therapeutics (https://doi.org/10.1016/B978-

0-323-99336-4.00018-5).

4.Sujata Prasad, Amar Singh, Shuxian Hu, Wen Sheng, Priyanka Chauhan, James R Lokensgard (2023) Dysregulated brain regulatory T-cells fail to control reactive gliosis following repeated antigen stimulation. iScience DOI: 10.1016/j.isci.2023.106628

5.Sujata Prasad, Shuxian Hu, Wen Sheng, Priyanka Chauhan, James R Lokensgard (2021) Dysregulated microglial cell activation and proliferation following repeated antigen stimulation.

Frontiers in Cellular Neuroscience 2021 doi: 10.3389/fncel.2021.686340

6.Priyanka Chauhan, Wen Sheng, Shuxian Hu, Sujata Prasad, James R Lokensgard (2021) Differential cytokine-induced responses of polarized microglia. Brain Sci Nov 10. doi:

10.3390/brainsci11111482.

7.Priyanka Chauhan, Shuxian Hu, Wen Sheng, Sujata Prasad, James R Lokensgard (2020) Programmed death ligand-1 induction restrains the cytotoxic T lymphocyte response against microglia. Glia Oct 31. doi: 10.1002/glia.23932.

8.Sujata Prasad, Shuxian Hu, Wen Sheng, Priyanka Chauhan, James R Lokensgard (2019) Recall responses from brain-resident memory CD8+ T-cells (bTRM) induce reactive gliosis. iScience 20 (512-526).

9.Amar Singh, Sujata Prasad (2019) The immune checkpoint protein programmed death 1 during HIV infection in children: clinical relevance and beyond. J Infect. Dis. 219 (9) 1353-1355 (Editorial)

10.Amar Singh, Sujata Prasad (2019) The quest for potential host blood biomarkers for tuberculosis diagnosis, severity, and treatment effects. EC Pulmonology and Respiratory Medicine 8.6: 2019: 484-486

11.Sujata Prasad and James R Lokensgard (2019) Brain-Resident T cells Following Viral Infection. Viral Immunology https://doi.org/10.1089/vim.2018.0084

Conferences/Talks/Posters (Total 23)

1.Invited speaker at virtual international conference on Healthcare Biotechnology, August 13 2021Organized by Center of Bioscience, Pillai College of Arts, commerce & Science, University of Mumbai, India.

2.Selected for Oral and Poster presentation, Investigators in training session at 17th International Symposium on Neurovirology: 04th June 2021 Virtual conference, USA

3.Oral presentation: Dysregulated microglial cell activation and proliferation following repeated antigen stimulation, Institute of Molecular Virology (IMV) seminar, December 9th 2020, University of Minnesota, USA.

4.Poster presentation: Recall response from brain-resident CD8 T-cells triggers iNOS production by microglia. Cold Spring Harbor meeting: Glia in Health & Disease (Virtual 2020).

5.Oral presentation: Recall responses from brain-resident memory CD8 T-cells (bTRM) trigger iNOS production by microglia, in Infection and Immunity Research Conference 10th March, 2019, department of Medicine, University of Minnesota, USA.

6.Invited speaker at International Conference on”Innovations and Translational Dimensions:

Food, Health and Environmental Biotechnology” [Biosangam- March 9-11, 2018] organized by Department of Biotechnology, Motilal Nehru National Institute of Technology, Allahabad, India.

7.Poster presentation: Recall responses from brain-resident memory CD8 T-cells (bTRM) induce reactive gliosis, in 16th International Symposium on Neurovirology, November 12-16, 2019, Atlanta, GA, USA.

Abstracts (Total 17)

1.Sujata Prasad, Shuxian Hu, Wen S Sheng, Priyanka Chauhan, James R Lokensgard. Dysregulated microglial cell activation and proliferation following repeated antigen stimulation, at 17th International Symposium on Neurovirology: 04th June 2021 Virtual conference, USA

2.Sujata Prasad, Shuxian Hu, Wen S Sheng, Priyanka Chauhan, James R Lokensgard. Recall response from brain-resident CD8 T-cells triggers iNOS production by microglia. Cold Spring Harbor meeting: Glia in Health & Disease (Virtual 2020)

3.Sujata Prasad, Shuxian Hu, Wen S Sheng, Priyanka Chauhan, James R Lokensgard. Recall responses from brain-resident memory CD8+T-cells (bTRM) induce reactive gliosis. J. Neurovirol. November 2019, volume 25 supplement 1, S35.

4.Sujata Prasad, Shuxian Hu, Wen S Sheng, Priyanka Chauhan, James R Lokensgard. Reactive glia promote development of CD103+CD69+ CD8+ T-cells through PD-L1. J. Neurovirol. April 2018 volume 13, supplement 1, S67.

5.Sujata Prasad, Shuxian Hu, Wen S Sheng, Priyanka Chauhan, James R Lokensgard. The PD-1: PD-L1 Pathway promotes development of CNS TRM cells following acute viral encephalitis. J. Neurovirol. 2016, volume 22, supplement 1, S62.

Extra-Curricular activities

1.Appointed as judge Twin Cities Regional Science Fair at Lee & Rose Warner Coliseum, Minnesota State Fairgrounds, Clough St & Judson Ave, St. Paul, MN 55108

2.Organized and presented poster in Society of Indian Human and Animal Mycologist, 2010

March in the Department of Ocular Microbiology, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.

3.Organized and attended CME on Mycotic Infections, April 2009 in the Department of Ocular Microbiology, Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.

References

1.Dr James R. Lokensgard

Professor

Department of Medicine

University of Minnesota

420 Delaware Street SE,

Email: ********@***.***

Minneapolis, MN 55455

2.Dr. Amar Singh, PhD

Assistant Professor

Department of Surgery

University of Minnesota

420 Delaware Street SE,

Minneapolis, MN 55455

Phone: 612-***-****

Email: ********@***.***

3. Dr. Wen Sheng (Collaborator)

Assistant Professor

Department of Medicine

Division of Infectious Disease & International Medicine

********@***.*** *************@*****.***

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