For detailed information on the current facility operations, please click here.
The mission of the Protein Technologies Facility (ProTech) is to further research in molecular and cell biology, protein biochemistry, and structural biology by overcoming major bottlenecks in these fields.
Our core services include molecular cloning, protein production and purification, and biophysical characterization of proteins. Since 2014, we also offer services surrounding the CRISPR/Cas9 genome engineering technology.
We offer customized generation of DNA constructs for recombinant protein production as a stand-alone service or as the first step in a protein production project. We are specialized in generating multi-gene expression constructs for production of protein complexes.
We offer recombinant protein production in E. coli, Drosophila S2 cells, insect cells using the baculovirus expression vector system (BEVS), and HEK293 cells.
Protein purification is offered as a full service at different scales. Due to limited capacity, for projects requiring purification of multiple proteins we also offer training and access to the equipment in our facility.
We offer protein characterization services using a variety of Biophysical techniques to study protein stability, oligomeric state, and secondary structure, or to analyze biomolecular interactions.
We also provide instrument training and access for some of these methods. To gain an overview of the techniques available, please see the Equipment section. More information can be found on the ProTech Wiki, accessible via MyVBCF. To discuss your biophysical characterization project, please contact Arthur Sedivy.
At this time, the Genome Engineering Facility is no longer able to accept new projects involving CRISPR/Cas9 mediated modifications of mammalian cell lines, generation of targeting constructs, or production of gRNAs. If you require help with plant genome engineering projects, please, contact Vera Schoft. For inquiries about the status of genome engineering projects initiated before July 2020 or the availability of reagents, please contact David Drechsel.
The OmniSEC system combines analytical size exclusion chromatography with right angle light scattering and measurement of refractive index, to accurately determine protein molecular weights and concentrations. It can be used to analyze the oligomeric state of proteins and protein complexes and to quantify the different species present in a sample. A full UV-vis spectrum and intrinsic viscosity measurements enable the accurate determination of extinction coefficients and shape/size of protein samples.
The PEAQ-ITC instrument can be used to perform isothermal titration calorimetry experiments to measure biomolecular interaction parameters. The change in enthalpy upon ligand binding is directly measured by the instrument and can then be used to determine the dissociation constant, stoichiometry, and other thermodynamic parameters of the binding interaction. Interactions in the high nanomolar to high micromolar range can be determined using the PEAQ-ITC.
The MASS-1 instrument is a system for measuring biomolecular interactions by Surface Plasmon Resonance (SPR). In addition to determining dissociation constants, SPR is ideal for measuring kinetics of interactions (kon and koff).
The SPR service is only possible as a full-service. In exceptional cases, access is offered to experienced users.
DLS is a label free technique that analyzes particle size distribution in a bulk sample. It is mainly used for protein quality control, as you can detect size distributions of molecules and therefore aggregation and oligomerization of your protein of interest. The Dynapro II plate reader can also be used for screening or long-term stability measurements. Experiments can be carried out for you; alternatively, you may receive training and can then book the instrument yourself.
Circular Dichroism is a spectroscopic technique that uses circularly polarized light to study the structure of chiral molecules, such as proteins. The CD spectrum of a protein can be used to determine protein secondary structure or some aspects of tertiary structure. CD can also be used to study protein stability using thermal melt analysis to determine protein melting temperatures (Tm). We perform CD on a Chirascan Plus CD spectrometer from Applied Photophysics. Experiments can be carried out for you; alternatively, you may receive training and can then book the instrument yourself.
MST is a novel technique used to determine molecular affinities. Thermophoretic changes upon molecular interaction can be monitored by fluorescent labeling or via tryptophan fluorescence, revealing binding constants in the range of nM to mM. Unlike many other methods, MST experiments can be performed rapidly and with small amounts of sample. We have two instruments for performing MST experiments, the NT.115 and the NT.LabelFree. MST experiments can be performed as a service, or you can receive training and then book the instruments yourself.
The NanoTemper Prometheus is a multi-purpose, label-free instrument measuring both intrinsic protein fluorescence and scattering while applying a defined temperature profile. Structural changes (folding/unfolding) as well as aggregation can be monitored over time, as a function of temperature, or under different chemical conditions. Since a sensitive fluorescence detection setup is used, only a minute protein sample is needed (>5µg/ml, 10µl). The Prometheus serves not only as a quality control instrument but also as a tool to find optimum conditions by using buffer screens to optimize stability. Using the Prometheus as a tool for day-to-day protein quality checks will improve the reproducibility of subsequent experiments that use purified proteins for structural and interaction studies while screening for potential binders can open up subsequent avenues of investigation.
The CreoptixTM WAVE is a novel, highly sensitive label-free biosensor used to analyze the affinity and kinetics of molecular interactions. The CreoptixTM WAVE provides similar information to the well-known Surface Plasmon Resonance biosensors but with increased sensitivity and lower noise, a superior “no-clog” microfluidics system allowing the detection and characterization of interactions in crude samples, and the ability to measure a very broad kinetic range (ka=103 – 3x109 M-1 sec-1; kd = 10-5 – 5 sec-1). Additionally, sample consumption, although dependent on the system studied, is very low. Combined with user friendly and robust analysis software, the CreoptixTM WAVE is ideal to study many different sample types and systems including those previously inaccessible using conventional biosensors.
If you are a new user, please contact David Drechsel (for ProTech core services), Arthur Sedivy (for biophysical characterization services) or Vera Schoft (for plant genome engineering services) to discuss your project requirements. To order services and book instruments, please use our web-based booking and request system. IMP, IMBA, GMI, and MFPL users can log in with their usual institute network accounts. External users must register. For questions, please contact protech(at)vbcf.ac.at.
Pricing information can be found on the ProTech MyVBCF Wiki site.
We require acknowledgement of facility use in publications.
A simple statement is sufficient and can be placed in the Materials and Methods section or in the Acknowledgments section, depending on the journal format.
The XXXXXX was performed by the Protein Technologies Facility at Vienna BioCenter Core Facilities (VBCF), member of the Vienna BioCenter (VBC), Austria.
In case of (co-)authorship:
The Vienna BioCenter Core Facilities (VBCF) Protein Technologies Facility acknowledges funding from the Austrian Federal Ministry of Education, Science & Research; and the City of Vienna.
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