Research Associate Analytical Development

JOHN PAUL KUNNATH

Boston, MA 857-***-**** *******.*@************.*** www.linkedin.com/in/john-kunnath EDUCATION

Northeastern University, Boston, MA Sep 2022 - Dec 2024 Masters in Biotechnology

SRM Institute of Science and Technology, Chennai, India Jun 2018 - May 2022 Bachelor of Technology in Biotechnology

WORK EXPERIENCE

Pledge Therapeutics, Research Associate – Virology, Canton, USA Present

• Maintain and passage multiple cell lines including RPTECs, HEK293T, Sf9 insect cells and Vero cells under sterile conditions using optimized split ratios to ensure consistent growth and viability

• Perform antiviral compound screening using BKV infection assays in RPTECs and analyze efficacy through qPCR and XTT-cell toxicity assays, processing 20+ plates weekly in a 386 well high-throughput format

• Harvest and propagate BK virus from Vero cells and prepare purified virus stocks for downstream infection assays

• Optimize virus amplification protocols, stock preparation and MOI calculations, achieving reproducibility with <10% variation in viral infectivity assays, ensuring reliable viral input for dose-response evaluations

• Collaborate with cross-functional teams in the US and EU to meet hit-to-lead screening milestones and SOP updates Solid Biosciences, Analytical Development Co-op – Molecular Biology, Charlestown, USA Jan 2024 – Jul 2024

• Conducted bridging studies for PCR systems to ensure data consistency across platforms and characterized AAV capsids for accurate quantification of capsid amounts

• Developed and implemented ddPCR (Singleplex/Multiplex) assays for high-throughput sample analysis, ensuring precise and accurate molecular measurement

• Led testing of over 400+ samples using the SpectraMax i3x platform and conducted Capsid ELISA for accurate AAV capsid characterization

• Collaborated on bioenergetic assays using the Octet system to optimize cell functionality and improve assay performance

• Created and validated protocols, delivering critical data that contributed to successful R&D and Process Development outcomes

Northeastern University, Application Lab RA, Boston, USA Sep 2023 - Dec 2023

• Cultured HEK-293 cells for transfection, maintenance and development studies, for optimizing cell culture conditions for optimized protein expression

• Performed CRISPR-Cas9 gene editing to knock out gene function, assessing the results through blue-white screening for accurate evaluation of gene disruption

• Operated the AKTA Pure 25 FPLC system for protein purification using Unicorn software to achieve chromatographic separation and other maintenance related things like priming pumps, buffer flushing

• Skilled in recombinant protein extraction and proficient in SDS-PAGE for assessing protein purity, molecular weight as well as western blotting

Takeda Pharmaceuticals, Advanced Analytical Development Intern, Cambridge, USA May 2023 - Aug 2023

• Designed and optimized bioenergetic assay kits used for studying cellular metabolism and energetics, assessing mitochondrial and glycolytic functions in real-time

• Performed mammalian cell culture on various CAR-T cell products, including media preparation and critical reagent transfers

• Screened 30+ donor samples under different conditions, such as post-thaw and post-culture, to evaluate cell metabolic performance and resilience

• Executed a comparison study between different assay kits using different donor screening processes to ensure accurate metabolic profiling and helped integrate assay results for product quality evaluation

• Successfully improved assay performance, and efficiency making it a tool for evaluating cell therapy products and supporting data-driven decisions and details

SKILLS

• Protein Biochemistry: SDS-PAGE, Western Blot, Sandwich ELISA, MALDI-TOF, Column Chromatography, MSD, CE-SDS, cIEF Cell Biology: Mammalian Cell culture, Transfection, Immunofluorescence, Cell Counting, Flow Cytometry, Cryopreservation Molecular Biology: qPCR, ddPCR, RT-PCR, Gel Electrophoresis, DNA/RNA Isolation, Nanodrop, Transformation, CRISPR-Cas9

• Technical- GraphPad PRISM, JMP Suite, Java, MySQL, AutoCAD, Microsoft Suite, ELNs (IDBS, Benchling) PUBLICATIONS

• Suresh V, Kunnath J, Reddy A. Prospective dietary radical scavengers: Boon in Pharmacokinetics, overcome insulin obstruction via signaling cascade for absorption during impediments in metabolic disorder like Diabetic Mellitus. Journal of Diabetes

& Metabolic Disorders, 1149-1169, 2022.https://doi.org/10.1007/s40200-022-01038-8

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