The Jagiellonian Center of Innovation provides a broad range of services in the field of proteomic analysis based on electrophoretic, chromatographic, and mass spectrometric techniques. The Center performs spectrometric analyses using Impact II QTOF system (Bruker), with an option of coupling to UHPLC chromatograph (Dionex). With this method, it is possible to separate complex mixtures of compounds and generate high-resolution mass spectra with mass measurement accuracy of 5 ppm (HRMS). The electrospray technique (ESI) is suitable for gentle ionization of studied molecules, while fragmentation in the collision chamber enables identification of unknown compounds.
In the area of mass spectrometry, the Laboratory offers the following analyses:
- Determination of molecular weight of proteins/peptides
- Identification of proteins and peptides based on popular databases (UniProt, NCBI)
- Identification and localization of post-translational modifications (PTMs)
- Label-free protein quantification.
The proteomic services listed above are complemented by protein characterization by electrophoretic, immunoenzymatic, and physicochemical methods. A wide array of techniques allows for the separation of proteins in complex mixtures, identification based on their molecular weight, physicochemical characterization, and analysis of a selected protein. The Laboratory also performs protein purification with the use of low pressure chromatography.
We offer a service of isolation and analysis of proteins from complex mixtures (e.g. milk, body fluids). The laboratory has the ability to purify proteins from various sources. We conduct research and development projects in the field of isolation and biochemical analysis of proteins.
A number of methods are available in the laboratory that allow for biochemical characterization of proteins:
- Gel electrophoresis (SDS-PAGE, Native-PAGE) – Mini-PROTEAN Tetra Cell (Bio Rad)
- Western Blot – Mini Trans-Blot® Cell (Bio Rad)
- Multiangle light scattering (MALS) coupled with chromatographic system – DAWN8+ detector (WYATT) – protein molecular weight analysis in solution
- UV-VIS spectroscopy – Evolution 201 (Thermo Scientific)
- Microscale thermophoresis (MST) – Monolith NT.115 (Nanotemper) –
- Analysis of protein interactions Surface plasmon resonance (SPR) – Biacore X-100 (GE Healthcare) – analysis of protein complex binding kinetics Enzyme immunoassay
- (ELISA) – qualitative/quantitative determination of protein using immunoenzymatic assay
The laboratory has experience in conducting research and development studies in the field of biopharmaceuticals. Our team prepares the results for scientific publications. We offer substantive and practical support for start-ups in proteomic research.