Markers of isoelectric point – an essential tool in capillary isoelectric focusing

0584027 - ÚIACH 2025 DE eng A - Abstract
Duša, Filip - Dadajová, Pavlína - Čmelík, Richard - Moravcová, Dana - Šlais, Karel
Markers of isoelectric point – an essential tool in capillary isoelectric focusing.
CE Forum 2024. Aalen: Aalen University, 2024. s. 14-14.
[CE Forum 2024. 28.02.2024-29.02.2024, Aalen]
R&D Projects: GA ČR(CZ) GA22-00236S
Institutional support: RVO:68081715
Keywords : capillary isoelectric focusing * protein separation * pI markers
OECD category: Analytical chemistry

Capillary isoelectric focusing (cIEF) is a powerful technique for separating proteins, peptides, and even bioparticles. However, it is essentially dependent on access to a robust set of internal standards (pI markers) to determine of the isoelectric points (pI) of the analytes. Development and characterization of internal pI markers based on derivatization of low-molecular-mass molecule cores highly absorbing in UV-vis spectra to provide a set of unique ampholyte structures. Each of the used cores gave the derivatized structures unique absorbance spectra. Using multi-wavelength detection, the pI markers could be selectively identified and the pI values of the analytes precisely determined. Careful calibration of the pI of the developed structures was crucial and an established cIEF protocol enabled precise pI determination of the developed structures. More than one hundred selectively detectable pI markers for cIEF and other formats of isoelectric focusing have been developed and are presented in this work.
Permanent Link: https://hdl.handle.net/11104/0352028