Research & Development
Specialists in Recombinant DNA products, our expertise extends over –
- Cloning of Genes in various expression systems
- Process optimization and scale-up of Protein Expression
- Downstream Processing (Protein purification, Protocol Development and Characterization of Proteins)
- Fermentation Technology
- Tissue Culture Assays
- Contract Research Services
Efficient production of recombinant proteins mean relying on intelligent protein expression systems. This becomes very important if the protein is to be expressed using an active ingredient of human or animal. At Sudarshan Biotech plasmids are designed for high expression levels of target proteins and on request, tailor made expression systems can be developed.
Sudarshan Biotech also addresses the following critical factors during bacterial fermentation like: media composition, oxygen supply, feeding strategy, time of induction, growth rate, by-products, product variants etc. Based on these parameters, robust and scalable fermentation protocols resulting in high product yields are developed which are readily transferable to contract manufacturers working according to GMP standards. At Sudarshan Biotech, fermentation equipments are designed upto a scale of 50-100 liters culture volume, which allows production of several kilograms biomass for subsequent recovery of the target proteins.
Protein Purification
Purification of proteins can be done from various sources like: E. coli, Pichia Pastoris and Mammalian cells. Cell disruption, purification, and polishing steps are also done. For this purpose we choose the appropriate purification methods/principles to remove all undesired contaminants such as host cell protein (HCP) and pyrogens / endotoxins and DNA. Precipitation and filtration methods, affinity, ionic exchange, hydrophobic interaction, hydroxyl apatite and size-exclusion chromatography etc. are applied. Optimized reports and necessary documentation are given as they are mandatory for approvals by regulatory authorities.
Seven Stages Of Protein Process Development
The development of a production process can be divided into at least seven different stages:
- Genetic engineering using expression vectors
- Optimization and characterization of host vector systems
- Fermentation process optimization and scale up
- Down stream processing
- Product and process related analysis
- Scale up
- GMP Process transfer
Physicochemical Analysis
- Electrophoresis (SDS-PAGE, Western Blot)
- HPLC (reversed phase, size exclusion, ion exchange)
- Spectroscopy (UV-VIS, fluorescence, circular dicroism)
- N-terminal sequencing
Biochemical Analysis
- Determination of DNA contamination and host cell impurities (Threshold™ system)
- Determination of the endotoxin content
- Enzyme assays to determine and quantify the catalytic activity of the target protein
- Determination of the protein content and activity by ELISA
Adding power to our research labs are the latest systems like:
Ultracentrifuge; -80 C freezer and -20 freezer; PCR machines; Sonicators; Incubator Shakers; Lab Scale fermentors; Gel Documentation system; DNA sequencer; Milli-Q- Water purification unit; Lyophilizer; Several Laminar flow-hoods, Walk-in cold room; Fully Air conditioned – 37.5 Tons Generator – 125 KVA – 3 seconds backup for sensitive equipment; UPS-10KVA