Quantitative analysis of peptides/proteins

Quantitative analysis performed in the Laboratory allows either to measure the absolute amount of a particular protein, or to compare the protein levels in two or more groups of biological samples. For the relative analysis, both targeted (hypothesis-driven) and global (shotgun) approach is available.

Global relative proteomic analysis - is performed for complex protein mixtures (body fluids, cell compartments etc.) on the Orbitrap Velos mass spectrometers. Methods requiring stable isotope labeling of samples (iTRAQ, SILAC), as well as label-free approaches, can be used for this type of quantification and are selected individually in accordance with experiment design. For each experiment, data validation and statistical analysis is performed, to obtain a list of proteins which show a significant abundance change between control and experimental samples. The global relative analysis is mainly used in differential proteomics experiments.

Targeted analysis - performed on Waters Xevo instruments, working in the MRM (multiple reaction monitoring) mode. This type of analysis provides selective quantitative information on the proteins of interest and is less time-consuming than global analysis of a whole proteome. Targeted analysis, fast and suitable for routine measurements, can be easily adapted for screening or diagnostic purposes.

The Laboratory also offers measurements of absolute protein concentration with AQUA (AQUA - absolute quantification of proteins) strategy. This type of analysis is performed on Waters Xevo mass spectrometers and requires synthetic peptide standards, which can be prepared by the facility cooperating with the Lab.

The quantitative analysis of proteins/peptides is not a routine service. Please contant the Head of the Laboratory, Prof. Michal Dadlez (This email address is being protected from spambots. You need JavaScript enabled to view it.) for a detailed information on costs and conditions. The staff is always ready to give advice and information on sample preparation procedures and interpretation of results.

Quantitative analysis of low molecular weight compounds

The analysis of low molecular weight compounds in complex matrices (mainly biological, i.e. body fluids) is performed on the triple quadrupole mass spectrometers (Waters Xevo TQ oraz Waters Xevo TQ-S) working in the MRM mode. Various ionization techniques (ESI, APCI, APPI, MALDI) can be used, if needed. The Laboratory is ready to develop a novel quantitative method for a compound of choice, or to adapt a known analytical approach for routine use. Measurements of this type are suitable for screenings, clinical diagnostics or drug monitoring, both in pre-clinical and clinical phase.

Costs and conditions of analysis can be discussed with the Head of the Laboratory, Prof. Michal Dadlez (This email address is being protected from spambots. You need JavaScript enabled to view it.). For additional information concerning technical issues, please contact the staff.