RAGUNATH CHANDRAN

Highly accomplished Protein Production / Protein Biochemist Leader and troubleshooting specialist. I have extensive and diverse experience in purifying a myriad of proteins using different expression systems including but not limited to mammalian cells, insect cells (sf9,sf21&Tni), BacMam technology, E. coli, yeast, in vitro translation, and plant system.

• Successfully purified single and complex proteins in the following companies: X-Chem, ZebiAI, Relay, Roivant, and EXO Therapeutics.
• Specialized in protein engineering and purification, proven track record in protein purification by using AKTA, as well as large-scale protein purification (with and without tag) using ion-exchange (IEX), affinity and size exclusion chromatography (SEC).
• in vitro assays, radioactive and fluorescent labeling technologies, protein crystallization, cell-based assay development.
• Proactively forecast and manage capacity in anticipation of rapid portfolio growth, fluidly adapt to increase in demand by building out, improving efficiency, and judiciously utilizing CRO efforts.
• Motivated hard worker, quick learner, team-oriented, and problem-solving capability.
• Interact with staff of other departments to coordinate information and policies and procedures.

Biochemistry:

• Protein expression from mammalian cells (up to 50 L scale), insect cells (Sf9) (up to 75 L scale), E. coli (up to 25 L scale), and plant system and purification using AKTA Pure 25.
• Protein purification by ion-exchange (IEX), affinity and size exclusion chromatography (SEC), tangential flow filtration (TFF), ultrafiltration / diafiltration (UF/DF), purification of inclusion body and protein refolding, isolation of membrane protein, protein estimation using different techniques, aSEC, SEC-MALS, Fluorescence Polarization, TR-FRET, DSF, or Alpha Screen.
• in vitro enzyme kinetics, inhibitor study, assay development, in vitro protein labeling and protein crystallization. ELISA, IEF, PAGE gel (SDS & Native) electrophoresis (1D and 2D) and western blotting.
• Protein-Protein interaction, UV/Vis, and Luminescence Spectroscopy. Polysaccharide purification and estimation by colorimetric method, HPLC, and TLC.

Molecular Biology:

• DNA and mRNA isolation, purification, and estimation.
• PCR, RT-PCR, Q-PCR, site directed mutagenesis, vector construction.
• In vitro recombination and Gateway vector cloning. Southern and Northern blotting.

Microbiology:

• Culturing aerobic and anaerobic bacteria (Pectobacterium Carotovorum, Aggregatibacter actinomycetemcomitans, Actinobacillus pleuropneumoniae, Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus gordonii, Streptococcus mutans, Fusobacterium nucleatum, Bordetella bronchiseptica and Pseudomonas aeruginosa), BLS-2, cell viability assay.
• Transposon mutagenesis and genetic complementation. Bacterial biofilm culture in tissue culture plates, polystyrene rods, intravenous catheters, assay for autoaggregation and biofilm detachment by crystal violet staining, FISH technique, live /dead cell assay, immunofluorescence, cell-based assay.
• Single/Multiple bacterial biofilm formation. Display technology, immunohistochemistry.

Eukaryotic Cell Culture:

• Culture Spodoptera frugiperda (Sf9, sf21 & Tni), HEK 293F & Expi 293, CHO, HeLa Cells, Human Buccal Epithelial and Fibroblast cells.

Plant Molecular Biology:

• Plant transformation using Agrobacterium and particle gun gene delivery system. Plant tissue culture, micropropagation, and acclimatization of transgenic plants.
• Protein extraction, estimation, recombinant protein purification, and in vitro enzyme assay.

Virology:

• Preparation, purification, and infection of baculovirus and BacMam system

Macromolecular Crystallography:

• Crystal tray setup (both manual and using robot), screening, and optimization.

Analytical Techniques:

• AKTA Pure 25 (Unicorn 7.3) and VARIAN-HPLC chromatographic method development.
• UV and Mass spectroscopy, ion-exchange, affinity, and size exclusion chromatography.
• Experience with method development and validation and be familiar with GLP/GMP practice and SOPs, assist in the processing/reviewing of all specifications related to protein purification and documentation as needed.
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Leadership:

• Mentored Postdoctoral fellows, Graduate students, and protein production team to help in designing experiments, expression, and purification of proteins, analyzing data, writing reports, and presenting data at seminars and conferences.

Technical Skills:

• MS office, Photoshop, and Bioinformatics tools.

• Transgenic plants expressing dispersinB, Publication number: 20140020138.
• Recombinant transfer vectors for protein expression in insect and mammalian cells, Publication number: 20220251600.