research Experience

Internship Project
Process development for recombinant trypsin
Trypsin is a serine protease, having molecular weight near about 25 kD, secreted by acinar cells of pancreas as an inactive precursor trypsinogen. The activated trypsin cleaves the proteins at the carboxyl terminal end of accessible arginine (R) and lysine (K) amino acid residues. This property has been exploited by Biopharmaceutical industry for the production of insulin and insulin analogues like glargine, lispro etc. Activated trypsin cleaves proinsulin into insulin and C peptide. For the cost effective production of insulin and insulin analogues we have developed a process for the production of recombinant trypsinogen expressed in pichia pastoris. We have also optimized an activating mixture for the activation of trypsinogen. The developed activating mixture shows better activation of trypsinogen as compare to activating mixtures recommended by Sigma Aldrich. This In-house developed recombinant trypsinogen has a great impact on cost reduction at a full scale production.
Fermentation process development for therapeutic biomolecules
Improvements in process development and scalable fermentation technologies are increasingly important as biopharmaceutical industry progress from discovery to pilot and full scale production. In Biocon Ltd. I was involved in cost effective fermentation process development and scale up of insulin and insulin analogues like Glargine, Lispro, IN105 (oral insulin) and other biosimilars like HSA and hGH expressed in Pichia pastoris.
Place of work: RND/ PDP department, Biocon Ltd. Bangalore.
Instrument handled: HPLC (Agilent, dionex, waters) DIONEX ICS 3000, SKALAR (SAN ++)
Bioreactors (scale -10 Li, 2 Li)
Job profile
Scale up of fermentation technologies from seeding to production.
Optimization of bioreactor.
Documentation of experiments according to ISO specified standards

Dissertation Project
DNA Isolation and PCR optimization for RFLP and RAPD analysis of Jatropha.
Abstract: Currently Biodiesel industries are using jatropha curcus, jatropha integrita, jungly aerand etc. plants for production of biodiesel. Cultivation of all these plants requires large amount of land as well as time which intern increases the cost of the product. Therefore, it is rational to use highly efficient plant for the production of biodiesel. To find out the highly efficient plant for biodiesel production, DNA was isolated from all plants. Isolated DNA samples were digested by restriction enzymes and specific gene responsible for biodiesel production was amplified. It was found out that jatropha curcus species contains more number of specific genes for biodiesel production than other species and if use in biodiesel production can give higher throughput with minimum expenditure.
Place of work: Rajarshi shahu college, Latur, Maharashtra

Poster presentations
1) Presented poster on Correlation of cataract with collagen in National conference on Contribution of biological research to humankind held on 14th -15th December, 2007. Department of zoology, R. Bezonji Science College, Jalana, (Ms) India.
2) Presented poster on sericine activity of protein in National symposium on Emergence of Modern Techniques And Development In Biobusiness held on 18th-20th February 2008. Modern college of Arts Science and Commerce Ganeshkhind, Pune 53 (Ms) India.

Skills
Process optimization for therapeutic proteins at lab scale bioreactors.
Characterization and quantification of proteins.
Microbiological Techniques: Microscopy, Staining, Antibiotic assay, Isolation and Purification of specific strains, Enrichment technique.
Molecular Techniques: DNA isolation, Plasmid Isolation, Isoelectric focusing (IEF), ELISA, Western Blot, InVitro Transcription (IVT), RFLP, Gene cloning (Induction, over expression), PCR, mRNA Purification, AFLP, Enzyme Kinetics, DNA Fingerprinting, Restriction Mapping, Gel elution
Analytical Techniques: Agarose & SDS-PAGE, HPLC, UV Spectrophotometer, GC, TLC, Gel doc, protein estimation by Kjeldahls method.