Ashish Sharma, Ph.D. Email and Phone Number

■ Managed a high-performing team for biologics process development, focusing on novel antigen and antibody proof of concept and feasibility.■ Troubleshooting manufacturing challenges for several antigen and monoclonal antibodies (mAbs).■ Led the successful scale-up of antigen manufacturing, implementing rhamnose induction and conducting technical scouting and white space analysis.■ Established scale-down models for mAb-specific process development and optimized fed-batch strategies to attain high-titre antigen/mAb manufacturing..■ Designed strategies to convert transients to stable cell line, enabling consistent and high-yield monoclonal antibody production in mammalian cells.

Advisor: Joseph Shiloach, Ph.D. Senior Investigator. Biotechnology CoreResearch: Investigation of cellular responses in microbial and mammalian cell cultures expressing recombinant biologics. Identification of specific genes or pathways which are crucial to address the issue of low production of biologics in an bioreactor lab and industrial setting. Ascertaining the direct role of critical genes to the biologics manufacturing efficiency of the host-microbial and mammalian cells. ■ Developed a microarray and RNAseq data analysis pipeline to identify critical pathways and genes in mammalian (CHO, HEK) and microbial (E.coli, Pichia, Bacillus, yeast) host expression systems.■ Developed a process design for anthrax vaccine production (recombinant protective antigen) in non-sporogenic Bacillus anthracis strain. ■ Developed a unique microaerobic production process for over-expression of difficult to express toxin protein like toxA of C. difficile in E. coli.■ Investigated role of small RNA frnS in acid stress resistance during E.coli fermentation.■ Established and coordinated collaborative efforts to finish projects in a timely manner.■ Prepared initial drafts and performed subsequent revisions of all published work.■ Trained and Managed junior scientists.Technical Skills:Protein Expression and Purification ● RNA Purification ● cDNA libraries for RNA-Seq ● Assay Development and Optimization ● Transcriptome Analysis ● Primer Extension ● RT-qPCR ● Sterile Technique ● Basic Microbiological TechniquesBioinformatics Skills:Microarray and RNAseq data analysis Vector NTI, R Bioconductor, MeV tools, GeneSpring, PartekFlow, Partek genomics suite, CLC genomics workbench. Computer Skills:PC and Mac Operating Systems ● Microsoft Office ● Adobe Illustrator ● R Studio ● ImageJ ● Prism ● SigmaPlotAdministrative Skills:Project Management ● Science Communication (oral and written) ● Problem Solving ● Data Analysis and Processing ● Integrating Information ● Teamwork

Project: "RICEFUEL: Engineering enzymes, bacteria and bioconversion process for advanced Biofuels from waste grain straw".Advisor: Prof. K.J. Mukherjee, Dean, School of Biotechnology, JNU, New Delhi, India Dr. Syed Shams Yazdani, Group Leader- Microbial Engineering, Integrative Biology groupCoordinator- DBT-ICGEB Advanced Bioenergy Research, New Delhi, India■ Research task was to convert Rice Straw waste into fuel using the industrial microbe process organism, Geobacillus.■ Leveraged data and troubleshooting expertise to resolve obstacles in project progression.■ Established and coordinated collaborative efforts to finish projects in a timely manner.Technical skills:Process development for production of ethanol ● Systems and Synthetic biology ● Metabolic engineering ● DNA/RNA isolation ● Site-directed mutagenesis ● ELISA ● Fluorometry ● Quantitative PCR ● RT-PCR ● Gel electrophoresis ● Cloning.

Topic: Metabolic Engineering Strategies for Enhancing Recombinant Protein Expression in Escherichia coil.Advisor: Prof. K.J. Mukherjee, Dean, School of Biotechnology, JNU, New Delhi, India■ Managed multiple research project and utilized creative and critical thinking skills to address novel and complex questions.■ 1st Project: Generation of Quiescent cells using Indole assisted blockage of cell division in fed-batch fermentation.■ 2nd Project: Optimization of time of Induction and feeding strategy in the recombinant E.coli culture expressing Interferon beta,■ 3rd Project: Creation of a generalized platform for the over-expression of difficult to express recombinant proteins. Bioinformatic analysis of the time point microarray data was analyzed to identify critical genes which limit the recombinant protein production. Created 170+ single-gene deletion mutants and 45+ double gene deletion mutants in E.coli. Screened every single and double gene deletion mutant for the production efficiency of model protein eGFP and test protein L-asparaginase.■ Prepared initial drafts and performed subsequent revisions of all published work.■ Trained and Managed 4 junior scientists.Technical skills:● Recombinant protein expression ● Process development ● Cloning ● gene knock-in/knock-out ● Micro-fermentation (BioLector M2P labs) ● High throughput media optimization ● Clone screening in 48 well microtiter plate ● Scale-up ● Feeding strategies for optimal production ● Biostatistics (Design of Experiments, Regression analysis) ● Chromatography (HPLC-MS, GC-MS) ● RNA work (including library preparation for Microarray and RNA-seq)

Conducted a three-day workshop on--“Optimization and Scale-up of Recombinant Protein Production for Bacterial Systems” Organizing Agency: Biotechnology Industry Research Assistance Council (BIRAC) in collaboration with School of Biotechnology, Jawaharlal Nehru University from 22nd –24th August 2014■ Trained the scientists working on recombinant protein expression in 20 different biotech industries. ■ Gave lectures and hands-on training to participants on the applications high throughput screening technique for media optimization and protein overexpressing strain screening.■ Demonstrated use of DoE (Design of Experiments) software to decide the minimum, median and maximum value selection and Statistical analysis of Central Composite Design (CCD) for media optimization. ■ Provided hands-on practice on Janus Automated Liquid Handler and M2p BioLector microbioreactor for process development.

■ Developed high Throughput assay protocols in 48 and 96 well systems in the liquid handler (Perkin Elmer), implemented PAT (Process analytical technique) and QbD (Quality-by-design) approach for the production of Biosimilars/Biologics. ■ Extensively performed gene cloning, screening high expression clones, and process development in 48 and 96 well micro bioreactors (BioLector). Further screening for their sustained performance in high cell density batch, fed-batch, and chemostat fermentations. ■ Developed specific growth rate dependent feeding strategies for over-production of biologics in the best performing clones in fed-batch fermentations. ■ Developed an optimal fed-batch fermentation process to produce recombinant proteins (Interferon-β, Xylanase, and L-asparaginase) in E. coli. ■ Developed methods for quantification of intracellular and extracellular amino acids, carbohydrates, nucleotides in HPLC and GC-MS/MS. ■ Downstream process development, protein purification using AKTA chromatography, analysis with unicorn software.