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.
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 Cryocube F101h - deep temperature freezer
Eppendorf Innova 40 - benchtop small scale bacterial incubator
Anvajo fluidlab R-300 - portable photometer
IMAGE: Jörg Müller
IMAGE: Jörg Müller
Access & Team
Access
The facility is a fee-for-service (non-profit) technology platform operated by UKE. We provide support and expertise for local scientists, the international research community and users from industryaTrto the individual instruments is granted. Currently the biosafety training is given onsite.
Please get in touch with us to discuss your specific needs.
Please acknowledge any substantial contributions of the PP Facility to your research in publications. Please state the following in the acknowledgement section of your publication: “We acknowledge technical support by the PP facility at CSSB Hamburg”.
Please don't forget to let us know when you publish papers that acknowledge us. This will help us keep track and to justify the existence of the facility to funding bodies.
CSSB Centre for Structural Systems Biology
c/o Deutsches Elektronen-Synchrotron DESY
Notkestraße 85, Building 15
D-22607 Hamburg
Facility Operator: University Medical Center Hamburg-Eppendorf (UKE)
Publications
2025
Szal T, Petsolari E, Veliks J, Durante Cruz C, Paunina L, Madre M, Rachad F, Lewe P, Witt S, Tammela SP, Jirgensons A, Luisi BF, Windshügel B (2025) Disassembly of the Escherichia coli AcrABZ-TolC efflux pump by ligand-mediated disruption of TolC-AcrA interfacial contacts. bioRxiv. 638599; doi: https://doi.org/10.1101/2025.02.18.638599
2023
Schütz A, Bernhard F, Berrow N, Buyel JF, Ferreira-da-Silva F, Haustraete J, van den Heuvel J, Hoffmann JE, de Marco A, Peleg Y, Suppmann S, Unger T, Vanhoucke M, Witt S, Remans K (2023) A concise guide to choosing suitable gene expression systems for recombinant protein production. STAR Protoc. 4(4):102572. doi: 10.1016/j.xpro.2023.102572
Reichelt J, Sachs W, Frömbling S, Fehlert J, Studencka-Turski M, Betz A, Loreth D, Blume L, Witt S, Pohl S, Brand J, Czesla M, Knop J, Florea BI, Zielinski S, Sachs M, Hoxha E, Hermans-Borgmeyer I, Zahner G, Wiech T, Krüger E, Meyer-Schwesinger C. (2023) Non-functional ubiquitin C-terminal hydrolase L1 drives podocyte injury through impairing proteasomes in autoimmune glomerulonephritis. Nat Commun.14(1):2114. doi: 10.1038/s41467-023-37836-8.
Szal T, Chauhan SS, Lewe P, Rachad FZ, Madre M, Paunina L, Witt S, Parthasarathi R, Windshügel B (2023) Efflux Pump-Binding 4(3-Aminocyclobutyl)Pyrimidin-2-Amines Are Colloidal Aggregators. (2023) Biomolecules. 13(6):1000. doi: 10.3390/biom13061000.
Bodrenko IV, Zewdie TA, Wang J, Paul E, Witt S, Winterhalter M (2023) TolC-AcrA complex formation monitored by time dependent single-channel electrophysiology, Biochimie,0300-9084, https://doi.org/10.1016/j.biochi.2023.01.004
2022
Niebling S, Veith K, Vollmer B, Lizarrondo J, Burastero O, Schiller J, Struve García A, Lewe P, Seuring C, Witt S, García-Alai M (2022) Biophysical Screening Pipeline for Cryo-EM Grid Preparation of Membrane Proteins. Front. Mol. Biosci. 9:882288. doi: 10.3389/fmolb.2022.882288
Research
The PP facility is involved in the following research projects:
In-solution inhibition studies of SARS-CoV-2 3C-like main protease
As an extension of the crystallographic studies that Alke Meens at al. performed on DESY Campus, Cy Jeffries from the EMBL SAXS group (Dmitri Svergun) performs further extensive in-solution inhibition studies of the SARS-CoV-2 3C-like main protease using SAXS. The required proteins, including the SARS-CoV-2 3C-like main protease, is produced by the PPCF. CSSB Protein Production facility: Susanne Witt (UKE) Collaborators: Cy Jeffries (EMBL Hamburg)
Serological test for COVID 19 Description:
Our collaboration partner, the DDM (Diagnostics Development Laboratory) group at the BNI, has a longstanding expertise in the development of ELISA-based serological test being highly specific to distinguish between homologous viral subtypes. For the development of such subtype-specific Sars-Cov2 ELISA-based serological test the PPCF produces a variety of proteins and optimizes their formulation. CSSB Protein Production facility: Susanne Witt (UKE) Collaborators: Ben Rushton (DDM, BNITM)
