Microbiology support of protein recombinant expression

Access to banks of suitable expression systems, optimisation of production conditions and different purification protocols are used to produce recombinant proteins. Different E. coli and yeast expression systems are used to produce recombinant proteins and virus-like particles (VLP).

Three-module shakers Multitron II (Infors, Switzerland) are used to cultivate bacterial and yeast cells under controlled conditions in flasks.  Culture media is prepared using autoclaves Systec V-150 (Systec GmbH, Germany).

Bioreactor support

Twin 1 L EDF 1.1., 5 L BIO4 and 20 L BioFlo 410 Fermentors/Bioreactors are available for large-scale the cultivation of E. coli and yeasts.

Biomass collection and disruption

For analytical purposes, biomass is collected with Eppendorf analytical centrifuges. For larger quantities, Beckman J2-21 and AVANTI J-301 (Beckman Coulter, USA), Eppendorf 5810 R centrifuges are available.

The Soniprep 150, UD-20, UP200S ultrasound disintegrators, Microfluidizer LM 20 and Continuous flow cell disruptor CF1 are available for both analytical and preparative biomass disintegration.

Purification of proteins with chomatography

Laboratory scale chromatography systems ÄKTA prima plus, ÄKTA Avant (with Unicorn 6.2 software), and ÄKTA pure (GE Healthcare) are available for protein purification. The Unichromat 1500 machine is available for the work with thermolabile proteins.

Depending on the protein, different purification strategies are available: gel filtration on Sepharose CL2B, 4B, 6B, 4FF, 6FF, Superose 6, Superdex 75, Superdex 200 carriers, ion exchange chromatography using Sepharose Q HP, Fractogel DEW, Source 15Q and Cathionites Sepharose SP HP, Source 15SP. His-tag proteins use immobilized methal-affinity chromatography (IMAC). For special cases, complex purification protocols can be elaborated using hydrophobic interaction chromatography (HIC), reverse phase chromatography (PharmPrep HP 100 RP-18 and Lichroprep RP-18, 25-40 μm), as well as multi-modular chromatography carriers (hydroxyapatite).

Ultracentrifugation

Two ultracentrifuges L-100XP are available for purification of viruses and nanoparticles with sedimentation-based methods.

Process control and product characterization

A modular analytical prominence HPLC system LC20 (Shimadzu, USA) is used to control purification processes and characterize the purified product. Dynamic light diffusion (DLS) measurements for assessing particle size and protein homogeneity shall be performed with the Malvern Zetasizer Nano ZS equipment (Malvern instrument, UK).

Plant laboratory

Plant growth chambers MLR-351H and LGC300C, microbiological safety cabinet KR-130, Helios Gene Gun and Tubing Prep Station are available for plant growing and transfection experiments.

Anaerobic laboratory

PLAS LABS 855 anaerobic chamber is available for experiments requiring anaerobic conditions.