Medical University of South Carolina
Medicine
As Research Scientist, I led 3 highly collaborative projects focused on developing therapeutic alternatives for preventing kidney diseases using cell-based and animal disease models. Identify critical genes involved in the protein interaction network in kidney and evaluate how mutations associated with them are pathogenic at the cellular level.
I cloned, expressed and purified more than 10 proteins including full-length transmembrane proteins NEPH1, NEPHRIN using lentivirus and baculovirus production system in mammalian and insect cells respectively for biochemical & structural studies that helped in identification of a novel recovery mechanism induced by therapeutic protein HGF that aids podocyte cell recovery.
. I was responsible for designing experiments that drove the project forward towards submission 3 publications and one grant
. I Oversaw the cell based and animal studies and managed a small team of 3 research assistants.
As a Post doctoral Fellow at the Cell and Molecular Biology lab, in the Nephrology Division, at Medical University of South Carolina (August 2015 to December 2019), I
led 3 primary projects focused on studying mechanism of disease progression in kidney and developing novel assays to diagnose
kidney diseases employing cell based and biochemical platforms. I
developed a non-invasive, accurate and economical cell based reporter assay to detect an under-diagnosed form of a glomerular
disease (recurrent FSGS). CRISPR-Cas based knock-in cells were generated for amplification of the signal for reporter assay.
I used computational methods to identify novel small molecules that protects from injury-induced renal damage and validated them. These projects led to the submission of 15 publications, 1 patent and a postdoctoral grant to me and a STTR grant to group.
Institute of Microbial Technology, Chandigarh, India.
February 2009
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January 2015
Protein Science
During Ph.D., I carried out innovative research resulting in co- and lead-authored publications on structures of therapeutic and biological important proteins and their complexes. My work summary includes:
Analysis of the changes in the shape of the proteins (involved in host-pathogen interaction in HIV infection) unliganded and in complexes with their ligand:
• Global shape analysis of HIV-1 neutralizing antibody IgG1 b12, which confirmed that this neutralizing antibody is inherently rigid and is asymmetric in solution (BBRC, 2010)
• In pursuit of engineering mAbs for neutralization, employed structural and biochemical studies including SAXS, AUC, SEC-MALS, limited proteolysis, ELISA, Western blot analysis etc. to elucidate the differences in the shapes of 17 HIV-1 neutralizing and non-neutralizing mAbs. Complexes of mAbs with gp120 were also made and purified using HPLC, and SAXS data was collected and analyzed for complexes. Based on our results we have proposed a fresh look at our perception of mAb-mediated neutralization (JBC, 2014)
• Global structure analysis of tetravalent antibody CD4-IgG2 and its complexes with gp120 (dimeric to tetrameric complexes) (BBRC, 2011)
• Designed biosimilars to CD4 IgG2 and which currently in process of determining their neutralization efficacy with CD4 IgG2 and other neutralizing mAbs.
• Cloning, expression, and purification of cell surface glycoprotein CD4 (which interacts with envelope spike protein gp120) and generated domain deletion mutants and single point mutants of the linker region of CD4 to study the role of linker in CD4 mediated viral entry. ELISA, CD, SAXS, MALS and other biochemical and biophysical analysis confirmed that the native-like conformation of four domains CD4 is essential for gp120 mediated viral entry but it does not affect the binding of CD4 to gp120.
Vaccine design:
• Using KMP11, a Leishmania protein as a model system, we have shown that carrier protein influences immunodominance hierarchy which is an implication in vaccine design. Our result shows the presence of the epitope tag at the N and C terminal of native protein (KMP11) not only alters its global shape but also causes a difference in their immunogenicity (VACCINE, JUNE 2013).
Crystallization:
• I have ample exposure to X-ray crystallization technique as well and have 4 PDB submissions: 3SP3, 3RNX and 3RW8 and 3RT5 in RCSB.
DNA Protein Interaction:
• Molecular Docking Studies for HapR/DNA binding and confirming it by SAXS based study. Participated in visualizing the elusive open shape of G-actin in solution by SAXS data (JBC, 2011)
