Krupa Subramanian

• Served as the in vivo study lead for RNA platform projects focusing on LNP delivery of vaccine candidates to specific immune cells.
• Performed in vivo immunological assays characterizing vaccine- specific T-cell and B- cell subsets by intracellular cytokine staining (ICS) and multi-parametric flow cytometry (FACS) from mice and non-human primates (NHP)
• Acquired and analyzed multi-parametric flowcytometry data using DIVA, FlowJo, GraphPad (Prism) and Microsoft Office tools.
• Developed a combined Mouse B/Tfh Ex vivo flow panel to characterize the cellular response to vaccination in two months, providing pre-clinical data to support the commit to candidate data package for an important pipeline project.
• Drafted and submitted in-vivo study protocols, ensuring compliance with IACUC and regulatory standards.
• Managed in-vivo study logistics, including scheduling, study design, and coordinating with internal and external stakeholders.
• Authored comprehensive in vivo immunogenicity study reports for IND and Regulatory submission and patent filing and ensured all relevant documentation was submitted as per GSK data integrity policies
• Collaborated with study team to finalize third-party quotes, manage material shipments, and complete post-study processes.
• Assisted with onboarding new team members, training on in vivo study monitoring, drafting study protocols and reports, and routine in vivo work involving mouse spleen processing, flow staining and data analysis
• Served as the EHS representative for Immunology Department and was recognized with a bronze award for my contributions in 2023
• Received a silver award for my contributions as a successful in vivo study monitor on the GSK internal LNP candidate selection study

Promoted to Scientist 1 position

• Conducted HIV-1 quantitative viral outgrowth assays (QVOA) on clinical trial samples through a contract administered by the Division of AIDS (DAIDS) in the National Institutes for Allergy and Infectious Diseases (NIAID).
• Successfully developed and miniaturized the QVOA assay, a.k.a. the dQVOA, making it scalable and affordable.
• Second author in a publication on the dQVOA assay development work in PLos Pathogens (2019)
• Performed intracellular cytokine staining and dextramer staining.
• Acquired 12-18 color ex-vivo and intracellular flow cytometry independently using BD FACS Canto II or Stratedigm S1000Exi.
• Skilled in processing blood, bone marrow and tissue from humans and non-human primates with accuracy as per SOP.
• Experienced in primary cell enrichment through magnetic bead separation and DNA/RNA extractions using kits.
• Competent in culturing, expanding and assaying HIV-1.
• Received an Excellence award in 2018 for contributions at Southern Research Institute.

Lab relocated to CWRU after VGTI closure.

• Conducted HIV reservoir Assay quantification and studied the mTOR signaling in HIV patients
• Performed the TILDA assay (Tat/rev induced limiting dilution assay) to estimate the latent HIV reservoir size capable of reactivation in the CD4+ T cell compartment of virally suppressed subjects using qPCR to measure the production of HIV multi-spliced RNA.
• Quantified CD8 T cell killing of the reservoir in primary cells from viremic patients, using a cell-based flow cytometric assay to directly assess the cytolytic activity of antigen–specific CD8+ T cells.
• Optimized a flow-based autophagy panel to study inhibition of MTOR signaling pathway responses in CD8 T cells in different HIV patient types using dose response studies with Rapamycin and Torin.
• Investigated the HSF-1 siRNA-mediated downregulation of HSP70 expression and increased apoptotic levels in T-cells from elite controller HIV patients, using intracellular detection of heat shock protein expression by BD Phosflow and Annexin V staining techniques.
• Designed and optimized assays for the BioMark HD system from Fluidigm to enable the study of differential gene expression in high-throughput qPCR experiments at the 100-cell level in 96 and 384 well- plate formats.
• Sorted highly specific T-cell subsets for gene expression studies using microarray and next generation sequencing, and characterized subsets based on surface marker expression levels.
• Performed antibody-sandwich HIV-1 ELISA to detect soluble p24 antigen in culture supernatants.
• Validated gene expression at the single-cell level using the C1 platform from Fluidigm.

• Conducted high-throughput screening assay in 1536 format in BSL3 setting and managed the operation of liquid handling robotic systems.
• Performed molecular cloning and purification of DNA samples using mini- and maxiprep kits.
• Engaged in HIV work in BSL2+ and BSL3 settings, including large- scale production of concentrated recombinant and wild-type virus, titration, and quantitation.

• Isolated Mesenchymal Stem cells from Human Bone marrow, identified stem cell colony forming units, and performed immunocytochemistry and FACS analysis of the surface markers CD73 and CD90.