About
The Protein Production Core Facility (PPCF) is designed to provide users with the complete infrastructure for protein production using the most modern and efficient methods. Particular attention is paid to maintaining the highest quality standards of the samples to be produced, which guarantees the suitability of the samples for functional and structural studies in a reproducible manner. Additionally, we aim to develop sustainable processes.
Services
The Protein Production Core Facility offers all steps of protein production consisting of the mutagenesis and cloning of target genes in expression vectors, their heterologous expression in prokaryotic and eukaryotic hosts, as well as the subsequent purification of the corresponding proteins as a service. In addition to the full service, in which the employees of the facility carry out the work, the users can carry out the work in the facility under the supervision of the facility team. The service includes advice on the selection of suitable strategies, the provision and optimization of suitable protocols, the instruction and use of the required equipment and the provision of the required vectors, cell lines, chromatography materials and all necessary consumables. The following services are offered in detail:
- Cloning and mutagenesis
- Cultivation of prokaryotic and eukaryotic cells of risk group R2 or security level S2
- Transformation of prokaryotic cells
- Transfection of insect cells and mammalian cells
- High-throughput heterologous expression screening in plate format: E. coli & insect cells
- Heterologous expression in E. coli, insect cells and mammalian cells
- Cell harvest and cell lysis
- Ultracentrifugation and cell membrane isolation
- Protein purification using Äkta Pure chromatography devices
- Development and optimization of protein purification protocols
- Automation of protein purification protocols
- Biochemical characterization (SDS and native PAGE, IEF, Western Blotting)
- Biophysical characterization (stability measurements using microscale thermophoresis)
- E. coli strain collection
- Expression vector collection
- NEB freezer
Available Instruments
- Equipment for cloning and mutagenesis projects
- PerfectBlue Gelsystem Mini S, M, L
- Eppendorf Mastercycler X50a – PCR Cycler
- Bio-Rad ChemiDoc MP – imaging system
- Bio-Rad MicroPulser – electroporator for transformation of bacteria
- Bio-Rad GenePulser Xcell - electroporator for mammalian cells
- Infors HT Minitron – incubator for insect cell expression
- Eppendorf Innoa S44i – biological shaker for bacterial cell culture
- Eppendorf Innoa S41i – biological CO2 shaker for mammalian cell culture
- Infors HT Multritron - biological CO2 shaker for mammalian cell culture
- Bandelin Sonoplus - sonification equipment
- Microfluidics LM10 - cell lysing equipment
- Various Eppendorf Centrifuges 5920R - multi use centrifuge
- Eppendorf Centrifuge 5430R - refrigerated centrifuge
- Various Eppendorf Thermomixer C/F1.5
- 3x Thermo Fischer MaxiSafe 2020 – laminar flows
- 2x Beckman Coulter Avanti JXN-26 – harvesting centrifuges
- Beckman Coulter Optima Max-XP and Optima XPN-90 – ultracentrifuges
- GE Healthcare Äkta pure M - protein chromatography system
- GE Healthcare Äkta pure L - protein chromatography system, MicroKit and Shimadzu Fluorescence Detector available
- GE Healthcare Äkta Start - small scale protein chromatography system
- GE Healthcare Äkta FLUX S - Tangential Flow Filtration/Cross Flow Filtration
- Various BioRad power pacs (basic and HC)
- XCell SureLock Mini-Cell - PAGE system for self-made and ready-made gels
- Bio-Rad trans-Blot-Turbo - semi dry Western blotter
- Tecan SPARK 20M – plate reader
- Thermo Fischer Micromodulyo – freeze dryer
- Leica Fluorescence cell culture microscope
- Olympus CKX31SF – inverted cell culture microscope
- Eppendorf Cryocube F101h - deep temperature freezer
- Eppendorf Innova 40 - benchtop small scale bacterial incubator
- Anvajo fluidlab R-300 - portable photometer