Biotech Scientist bringing concepts with lab execution

• Large-scale purification of recombinant monoclonal antibodies secreted from mammalian host cells (client projects from international pharma companies)
• Trained and led colleagues within (5) and across teams (2) on bacterial protein and human intracellular protein expression and purification optimizations

Project: Understanding the structural role of membrane proteins in generating and maintaining the highly curved tubular morphology of the smooth endoplasmic reticulum (a phenotype disturbed in several neurodegenerative disorders)

• Cloned isoform-specific ORFs of five full-length integral transmembrane proteins (Protrudin, RTN2, RTN3, RTN4, Arl6IP1; size varying between 23 to 127 kDa) from mouse cDNA, designed and modified expression vector backbones
• Performed recombinant membrane protein expression in suspension HEK cells and large-scale membrane protein purifications for X-ray crystallography and cryo-electron microscopy
• Established suspension HEK cell culture in lab and corresponding transfection protocols
• Designed the project, conducted experiments, established new protocols and procedures, analyzed and interpreted results
• Trained internal and external lab students on the newly established protocols
• Established a new line of research in the lab and procured independent government research funding (ICMR-DHR) for this project

Project I: Biochemical and functional characterization of the cytoskeletal protein SEPT7

• Generated SEPT7 mutant constructs, established inducible mutant stable mouse embryonic fibroblast cell lines, and transiently transfected HEK and HeLa cells to study SEPT7 mutations by microscopic and biochemical assays
• Performed GTPase activity assays for recombinantly purified mutants from E. coli; data compiled in one publication and one review
• Trained technical staff (1)
• Developed and optimized parameters for an in-cell bioluminescent reporter assay to monitor SEPT7–SEPT7 and SEPT6–SEPT7 interactions in living adherent HEK cells in a dose-dependent manner

Project II: Production of subcomplexes of the ~1 MDa mammalian Apaf-1 apoptosome

• Subcloned human Apaf-1 ORF (137 kDa protein) into various expression vectors, designed truncation constructs for stable protein domains, and modified vector backbones
• Performed recombinant protein production in E. coli, Dictyostelium, and Sf9 insect cells; troubleshooted conditions for stabilizing aggregating protein sub-domains of Apaf-1
• Trained a user (1) and technical staff (1) on protein expression from Sf9 insect cells