Biological Analysis

The Jagiellonian Center of Innovation offers a wide range of analytical services in proteomics, based on electrophoretic, chromatographic and mass spectroscopy methods.

Electrophoretic analyses: biochemical analysis of proteins using electrophoretic methods, including SDS-PAGE 2D analysis, native electrophoresis and Western Blott.

Chromatographic analyses (WYATT)

In the proteomic laboratory of the Jagiellonian Center of Innovation, we can optimize and perform protein separations using SEC-MALS and SEC-MALS-QELS methods, with Heleos II and DynaPro NanoStar detectors. The detectors allow for determining mass and concentration without the necessity to use a reference. Moreover, with a DLS detector we can work in a cuvette mode and hence determine (at volumes of the order of several µl) various factors quickly, including:

  • relative and absolute molecular weight
  • determination of protein crystallization parameters
  • determination of protein melting point
  • measurement of conformation changes in the native environment (via change in hydration radius Rh parameter)

Mass spectrometry (QTOF)

The Jagiellonian Center of Innovation performs spectroscopic analyses using Impact II QTOF (Bruker) device, with optional connection to HPLC chromatograph. This allows for the generation of high-resolution mass spectra (HRMS) and low fragmentation of the studied molecules. The use of ProteinScape base and BioTools software (Bruker) included in the set enables:

  • automation of the spectra base search process
  • determination of protein structures
  • de novo protein sequencing along with optimization of results from very big matrices

Analysis of intermolecular interactions:

Based on MST and SPR methods using Monolith NT.115, Biacore X-100 or ELISA test, the Jagiellonian Center of Innovation offers analyses of intermolecular interactions, such as:

  • protein-protein interactions
  • protein-nucleic acid interactions
  • protein-lipid, protein-sugar interactions
  • protein-low molecular weight compounds interactions

MST (Monolith NT.115)

A method based on a microscale thermophoresis (MST) phenomenon that describes the mobility of molecules in a solution with a temperature gradient. This unique technique is based on the use of the phenomenon of molecule movement change in temperature gradient, which allows for:

  • analyzing protein interactions in solution
  • making the analysis independent of the chemical composition of the solution
  • the option to perform experiments in conditions similar to native ones


SPR (Biacore X-100)

The Surface Plasmon Resonance (SPR) phenomenon makes it possible to determine the strength of intermolecular interactions in real time. The SPR phenomenon is based on oscillation of conducting electrons present on the biosensor due to the incident light, which results in absorption of a strictly defined amount of the incident light wave, leading to a decrease in the energy of the reflected wave. The method also allows for:

  • determination of the studied analyte concentration
  • studying the affinity of the studied intermolecular interactions
  • analysis of kinetics and thermodynamics of the studied system
  • determination of association and dissociation constants of the complex formed upon the interaction

ELISA tests

This method is used for the detection of proteins in the studied material, using antibodies. Antibodies are conjugated with an enzyme that (upon the addition of the appropriate substrate) catalyzes the reaction of formation of a spectrophotometrically assayed product. The method is commonly used for scientific and diagnostic processes, such as:

  • determination of concentration and type of proteins in the studied sample
  • diagnostics of viral and bacterial presence
  • diagnostics of autoimmunological and neoplastic diseases

PCR, qPCR, RT-PCR

The Polymerase Chain Reaction (PCR) is a method that allows for the multiplication of specific DNA fragments in vitro. Real Time-PCR is a quantitative PCR reaction and it enables simultaneous multiplication of chosen DNA fragments and monitoring the amount of products formed in each cycle. The Jagiellonian Center of Innovation offers services within the scope of optimization of PCR and Real Time PCR reactions using a non-specific SYBR Green method or a specific method with complementary oligonucleotide probes, based on a fluorescence resonance energy transfer (FRET) between the molecules of a donor and a quenching agent. Additionally, we also offer RT-PCR (reverse transcription PCR), where RNA is used as a matrix.

The PCR technique has many applications mainly in genome studies, characterization of gene expression, gene cloning and clinical diagnostics.