Deepika Gautam

Accomplished Senior Research Associate experienced in designing and optimizing purification processes for vaccine candidates, including chromatography and tangential flow filtration. Developed scalable strategies and implemented regulatory-compliant viral clearance studies, enhancing product quality through collaboration with analytical teams. Demonstrated expertise in protein purification from various sources and advanced analytical techniques, contributing to efficient technology transfer and process characterization.

• CSIR-Institute of Genomics and Integrative Biology, New Delhi, India, Senior Research Associate, 04/24/24, 08/24/24
• Quantitative analysis of expression levels of various protein mutants relative to the wild type of protein was performed using flow cytometry (FACS) for in vivo cellular expression, and fluorescence spectroscopy for in vitro validation of fluorescence Intensity.
• The fluorescence ratio of GFP to mCherry was measured in cells co-expressing both reporters, enabling normalization and comparative assessment of folding efficiency and expression stability across different GFP mutant constructs., Protein mutants exhibiting significant deviations in fluorescence intensity compared to wild-type were subjected to purification via Ni-NTA affinity chromatography, facilitating downstream biochemical and structural characterizations of protein folding and stability properties.
• CSIR-Institute of Genomics and Integrative Biology, New Delhi, India, PhD scholar/Senior research fellow (SRF), 08/17/18, 04/24/24
• An acute pancreatitis model was established in Wister mice through intravenous administration of cerulenin, a known inhibitor of fatty acid synthase that induces pancreatic injury via disruption of lipid metabolism., Experimental animals received cerulenin, while control mice were administered physiological saline., At defined time points (6h, eight h,12h) post-injection, mice were euthanised, and a comprehensive tissue dissection was performed., Organs, including the pancreas, kidney, heart, and brain, were harvested for downstream analyses to assess the systemic and organ-specific effects of cerulenin-induced pancreatic injury.
• Quantification of damage-associated molecular patterns (DAMPs) such as mitochondrial DNA (mtDNA) and ATP was performed on serum samples from control and disease model mice by qPCR., Protein expression levels of cellular stress and apoptotic markers (Bax, Bad, caspases) were analysed in tissues from both the disease model and control mice using Western blotting.,

     Project: “Human Hsp60/10 interacts with dicarboxylic acids to mimic bacterial chaperone GroELES” 

• The chaperonin system in prokaryotes is GroELES. We have investigated the characteristics responsible for its chaperone function.
• • Cloning of GFP and its mutants into a pet-SUMO vector for purification and transformation into E. coli strains like BL-21(DE3), Rosetta, Stbl 2,3 cells.
• • Chaperones (GroELES, hHsp60/10, yHsp60/10, etc) were purified by using affinity chromatography (DEAE and ANX) and FPLC (AKTA pure and AKTA explorer)
• • GroELES makes it energetically favourable for a mutant GFP (K45E) to fold through a decrease in enthalpic barriers and reduces the entropic obstacles via a negatively charged environment within its cavity.

• AKTA Avant
• Explore
• PURE
• Flow cytometry (BD-LSR-II)
• Fluorescence spectroscopy (Fluorolog-3, Horiba)
• QPCR (Bio-Rad)
• PCR (Thermo)
• Microscope (confocal and Epifluorescence)
• Cryostat (Leica)
• Microtome (Leica)
• MST (Microscale thermophoresis)
• cell culture
• Microbiology
• Molecular biology
• Western blot
• qPCR