Ph.D Bioetechnolgy, Persuing Postdoc in cell and Molecular Biology

ASHISH KUMAR SOLANKI

Department of Medicine, Medical University of South Carolina, Charleston, SC, USA. E-mail **************@*****.*** ~ Mobile: +1-215-***-**** ********@****.***

CARREER OBJECTIVE

High potential Life Sciences Research Professional seeking challenging assignments in Life Sciences research in an organization of high repute where I can use my skills, find the opportunity to intensify my knowledge and aptitude; and offer me excellent working environment to help me and my group find way to progressive results.

SNAPSHOT

r PhD in Life Science on thesis entitled “Global structure analysis of monoclonal antibodies recognizing envelope components, gp120 and 41, of HIV-1” obtained from IMTECH-JNU PhD program.

r Masters of Science in Biotechnology from University of Jammu, through DBT Govt. of India in 2008.

r Bachelor of Science from Aligarh Muslim University, Aligarh (India) in 2005. TECHNICAL PREVIEW

Biophysical techniques:

Small/Wide angle X-ray scattering (SWAXS) Infrared Spectroscopy

X ray Crystallography ITC

HPLC, FPLC SEC MALS

Mass spectroscopic analysis Analytical Ultracentrifuge (AUC)

Ab initio modelling Circular Dichroism

Trained on solid and liquid phase peptide synthesis Molecular modelling, simulations docking

Molecular Biology and Recombinant DNA Techniques

Plasmid DNA isolation, Genomic DNA isolation from plants and microorganisms ̧ PCR, RFLP and RAPD analysis ̧ expertise in cloning and site directed mutagenesis ̧ DNA sequencing ̧ EMSA, RT- PCR, EMSA, Aptamer designing and amplification, ShRNA based gene knock down cell lines, CRISPR based gene knock out in cell lines

Protein: Expression and Biochemical characterization

Heterologous protein expression in bacteria, yeast and mammalian cell lines

Purification of recombinant proteins through various chromatographic techniques (like affinity, ion exchange, hydrophobic and gel exclusion chromatography)

SDS Electrophoresis

Immunoprecipitation and Western blotting

Working experience with glycoprotein

Experience of refolding of protein

Cell Culture Handling, Maintenance and Immunological Techniques:

Expertise in maintaining and working with various cell lines

Cell plating and preparation of transiently transfected as well as stable cell lines

Dual luciferase reporter assay for the detection of HIV-1 infectivity and antibody neutralization

FACS

ELISA

Immuno precipitation, Immunofluorescence, Confocal

Production and purification of polyclonal antibodies Hands on training on working with animals

Handled rabbits, Mice (FVB and BL-6), generation of knock out mice (inducible), Hens

(polyclonal IgY antibody generation for use in diagnostic kit) Microbiological Techniques/Running Bioreactor

Various techniques for culturing, growth, maintenance and preservation of bacteria Bioreactor operation for production of therapeutically important protein CD4 and gelsolin on large scale

Computational and Bioinformatics skills

Worked with Operating systems such as DOS and Windows

Knowledge of routine software as Microsoft Office, Adobe Photoshop, Endnote etc

Understanding of bioinformatics tools for day-to-day sequential analysis using BLAST and Clustal.

Primer designing using Gene Runner

Use of Graph-Pad Prism Sigma plot, Origin for statistical analysis of the data

Excellent training on model generation of protein based on Homology and fold recognition

Molecular dynamic simulation of peptide using Tinker

Molecular docking to understand protein-protein or protein-peptide interaction using

GRAMM and different docking servers, and their data analysis RESEARCH EXPERIENCE

PhD in the laboratory of Dr. Ashish Ganguli, Senior Scientist, Institute of Microbial Technology, Chandigarh, India

During my PhD, 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:

Experience in developing a diagnosis kit:

• Worked on project funded by GATES foundation entitled “accurate, accelerated and affordable kit to predict the pre-term and post-partum recovery”

• Experience of raised polyclonal IgY antibodies against a therapeutic protein “gelsolin”. Immobilized antigen on sepharose beads and from the pool of IgY, enriched (15%) the antibody against the antigen. Using the enriched antibody, we have developed a diagnosis kit to measure the level of this protein in human plasma. Also, I have worked on developing different affinity coating protocols including IgG, DNA aptamer based (screening and validation) and small molecule (peptide) based methods as well.

Structure analysis of proteins and their complexes involved in host-pathogen interaction in HIV infection:

• Global shape analysis of HIV-1 neutralizing antibody IgG1 b12, which confirmed that this neutralizing antibody is inherently rigid and is asymmetric in solution.

• 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 nonneutralizing mAbs. Complexes of mAbs with gp120 were also made and purified using HPLC, and SAXS data was analyzed for complexes. Based on our results we have proposed a fresh look at our perception of mAb-mediated neutralization.

• Global structure analysis of tetravalent antibody CD4-IgG2 and its complexes with gp120

(dimeric to tetrameric complexes).

• Designed biosimilars to CD4 IgG2 and currently comparing 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 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. Crystallization:

• I have ample exposure of 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.

POST-DOCTORAL RESEARCH EXPERIENCE:

Interaction studies of slit diaphragm protein in kidney: I have a large amount of experience in the field of protein biochemistry and have gained a substantial knowledge of podocyte biology during the years spent as postdoc in Dr. Deepak’s lab working on podocyte biology. I have cloned and purified several slit diaphragm proteins and their interacting partners including Neph1, Nephron, Myo1, Zo-1 etc. These genes were cloned in lentiviral vector pBABE puro, and then using Phoenix retrovirus producer cell line for the generation of helper-free ecotropic and amphotropic retroviruses, generated stable cell lines (HEK and human podocytes cell lines) expressing these proteins. Purified these proteins and pulled down were performed to decipher their interaction network Additionally, I have a postdoctoral grant for working in a project where we have identified a novel mutation in the gene CLCN5 which codes for an endosomal chloride/hydrogen exchanger in a family with an X-linked form of focal segmental glomerulosclerosis. The goal of this project is to understand the mechanism(s) by which this mutation is pathogenic at the cellular level. This interesting finding of a mutation in a renal tubular protein as the cause of the glomerular disease raises the possibility that understanding how this gene variant causes glomerular injury may help understand a much wider spectrum of chronic kidney disease. This project will be an ideal platform for me to move into translational biology where I can correlate the clinical data with basic sciences to enhance human health and well-being.

PUBLICATIONS

Published:

1. Global structure of HIV-1 neutralizing antibody IgG1 b12 is asymmetric. Ashish, Solanki AK, Boone CD, Krueger JK, (BBRC, 2009) 2. SAXS data analysis and modeling of tetravalent neutralizing antibody CD4-IgG2 -/+ HIV-1 gp120 revealed that first two gp120 bind to the same Fab arm. Rathore YS, Solanki AK, Dhoke RR, Ashish., (BBRC, 2011) 3. Evidence on how a conserved glycine in the hinge region of Hapr regulates its dna binding ability: lessons from a natural variant”.

Dongre M, Singh NS, Dureja C, Peddada N, Solanki AK, Ashish, Raychaudhuri S.( JBC, 2011) 4. Visualizing the elusive open shape of G-actin in solution by SAXS data analysis Sagar A, Peddada N, Solanki AK, Choudhary Vikas, Garg, R., Ashish (BBRC May, 2013) 5. Carrier protein influences immunodominance hierarchy: implication in vaccine design. Moumita Ghosh, Ashish K. Solanki, Koushik Roy, Reema R. Dhoke, Ashish and Syamal Roy

(VACCINE, JUNE 2013)

6. Global shape and ligand binding efficiency of the HIV-1 neutralizing antibodies differs from the ones which cannot neutralize.

Solanki AK, Rathore YS, Badmalia MB, Dhoke RR, Nath SK, Nihalani N and Ashish (JBC, October, 2014)

7. Adriamycin susceptibility among C57BL/6 substrains. Ehtesham Arif, Solanki AK and Deepak Nihalani (Kidney International, 03/2016) 8. Structural analysis of the Myo1c and Neph1 complex provides insight into the intracellular movement of Neph1

Arif E, Sharma P, Solanki AK, Mallik L, Rathore YS, Twal WO, Nath SK, Gandhi D, Holzman LB, Ostap M, Ashish, Nihalani D (Mol Cell Biol., 2016) AWARDS AND FELLOWSHIPS

Qualified for Junior Research Fellowship from CSIR-NET in 2007 and 2008

Qualified Graduate Aptitude Test in Engineering (GATE, 2008).

Qualified for Jawaharlal Nehru Fellowship for master program with All India Rank 194.

Selected as Junior Research Fellowship (JRF) and Senior Research Fellowship (SRF) at Institute of Microbial Technology in Chandigarh in Feb 2009 and 2011 respectively.

Recipient of the prestigious Ben J Lipps Research Fellowship funded by American Society of Nephrology for the period of 2016-2018

Invited reviewer for journal “Frontiers in Immunology”

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