Naslov
Determination of isoelectric points for pI markers in capillary isoelectric focusing

Autori
Ansorge, Martin ; Šteflova, Jana ; Pocrnić, Marijana ; Moldovan, Alina ; Vigh, Gyula

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, ostalo

Izvornik
18th International Symposium and Summer School on Bioanalysis, Book of Abstracts / Kilar, Ferenc - Komarno : J. Selye University, 2018, 48-48

ISBN
978-80-8122-241-2

Skup
18th International Symposium and Summer School on Bioanalysis

Mjesto i datum
Komárno, Slovačka, 25.06.2018. - 30.06.2018

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
capillary isoelectric focusing, dissociation constant, isoelectric point

Sažetak
Capillary isoelectric focusing is an electromigration analytical method. As it is, it serves mainly for separation of ampholytic compounds, such as proteins. Therefore, it finds great application in the biopharmaceutical industry. In this technique, species separate according to their isoelectric points in a stable linear pH gradient in separation capillary. Since commercial gradient mixtures consist of up to several hundreds of ampholytic components (so called carrier ampholytes), forming the pH gradient is a time demanding procedure. The formation of the pH gradient can be visualized by introducing so-called pI markers into the carrier ampholytes. These pI markers should be well-defined small ampholytic molecules of good focusing abilities. This means that their - dz/dpH around the isoelectric point is as high as possible to achieve sufficient focusing power. This work focuses on determination of dissociation constants and isoelectric points of various, commercially available compounds, which can serve as pI markers for cIEF. Since thermodynamic dissociation constants are not tabulated and values provided in literature can vary according to the determination method, here Archem SPARC and ACD Lab software is used to estimate pKa values of each potential candidate. For compounds that fulfilled our requirements we measured their electrophoretic mobilities within a pH range of 2 – 12 and determined the dissociation constants and isoelectric points.

Izvorni jezik
Engleski

Znanstvena područja
Kemija

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