BIOCentre’s Microbiology lab specializes in two types of services – protein production and training. Microbiology lab performs expression (µg to mg scale), purification and process development of recombinant proteins, biopharmaceuticals and biosimilars. Microbiology lab offers hands-on-training courses on separate and parallel bioreactor systems (BIOSTAT Qplus 12 x 0.5L, B 2 x 2L, Cplus 30L), various cloning and general microbiology lab techniques. We also provide bacteria and yeast cell banking.
BACTERIAL CELL TRANSFORMATION
Transforming competent E.coli cells by heat shock with an expression vector carrying the target DNA, encoding the protein of interest. Selection of the transformed E.coli on selective agar plates and preparation of glycerol stocks for further use.
PLASMID PROPAGATION AND PURIFICATION
The competent E.coli cells are transformed with the desired template DNA provided by customer. Extracted DNA prep (mini, midi or maxi) quality control includes separation on agarose gel, A260/280 ratio, and quantification.
PROTEIN EXPRESSION IN E. COLI OR YEAST
Recombinant protein expression involves cultivation of transformed cells in desired conditions and scale: 1L shaker flask, 0,5L fermenter (up to 12 in parallel), 2L fermenter (up to 2 in parallel), or 30L fermenter. Preceding the large scale production, we offer an analytical (small scale of 100mL) expression of recombinant protein. To maximize the content of soluble recombinant proteins we optimize the expression conditions including choice of medium, carbon source, temperature, pH, aeration, inducer concentrations, and length of induction.
PROTEIN DOWNSTREAM PROCESSING is done in our DOWNSTREAM PROCESSING LAB
Downstream processes include cell lysis (high pressure and ultrasonic homogenizers), protein separation and purification. Desalting, buffer exchange and sample concentration can also be done.
PROTEIN QUALITY CONTROL is done in our BIOANALYTICAL PROTEOMICS LAB
SDS-PAGE and western blot analysis of the recombinant protein samples from analytical or production scale. NanoDrop quantitation, LC-MS/MS peptide mapping and protein identification.
DNA, RNA and protein extractions from microbial cells. Protein SDS-PAGE and western blotting. PCR detection and identification of microorganisms in different samples e.g. probiotics, beer spoilers, and other food and beverages.
EQUIPMENT AND INSTRUMENTATION
Sartorius BIOSTAT Qplus 12 fold 0.5L MO, BIOSTAT B Twin 2L MO, BIOSTAT Cplus 30L MO; high pressure homogenizer Panda Plus, Geo Niro Savi; ultrasonic homogenizer Bandelin, Sonoplus; NanoDrop One, Thermo Scientific; Sorval Stratos continuous flow centrifuge.