Sampling strategies for capillary isoelectric focusing with electroosmotic zone mobilization assessed by high-resolution dynamic computer simulation

Takácsi-Nagy, Anna; Kilár, Ferenc; Páger, Csilla; Mosher, Richard A.; Thormann, Wolfgang (2012). Sampling strategies for capillary isoelectric focusing with electroosmotic zone mobilization assessed by high-resolution dynamic computer simulation. Electrophoresis, 33(6), pp. 970-980. Wiley-VCH 10.1002/elps.201100525

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The impact of initial sample distribution on separation and focusing of analytes in a pH 3–11 gradient formed by 101 biprotic carrier ampholytes under concomitant electroosmotic displacement was studied by dynamic high-resolution computer simulation. Data obtained with application of the analytes mixed with the carrier ampholytes (as is customarily done), as a short zone within the initial carrier ampholyte zone, sandwiched between zones of carrier ampholytes, or introduced before or after the initial carrier ampholyte zone were compared. With sampling as a short zone within or adjacent to the carrier ampholytes, separation and focusing of analytes is shown to proceed as a cationic, anionic, or mixed process and separation of the analytes is predicted to be much faster than the separation of the carrier components. Thus, after the initial separation, analytes continue to separate and eventually reach their focusing locations. This is different to the double-peak approach to equilibrium that takes place when analytes and carrier ampholytes are applied as a homogenous mixture. Simulation data reveal that sample application between two zones of carrier ampholytes results in the formation of a pH gradient disturbance as the concentration of the carrier ampholytes within the fluid element initially occupied by the sample will be lower compared to the other parts of the gradient. As a consequence thereof, the properties of this region are sample matrix dependent, the pH gradient is flatter, and the region is likely to represent a conductance gap (hot spot). Simulation data suggest that sample placed at the anodic side or at the anodic end of the initial carrier ampholyte zone are the favorable configurations for capillary isoelectric focusing with electroosmotic zone mobilization.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Service Sector > Institute for Infectious Diseases > Laboratory for Clinical Pharmacology

UniBE Contributor:

Thormann, Wolfgang

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0173-0835

Publisher:

Wiley-VCH

Language:

English

Submitter:

Wolfgang Thormann

Date Deposited:

04 Jul 2014 09:28

Last Modified:

22 Dec 2014 11:06

Publisher DOI:

10.1002/elps.201100525

PubMed ID:

22655305

Uncontrolled Keywords:

Carrier ampholytes, Electroosmosis, Isoelectric focusing, Sample application, Simulation

BORIS DOI:

10.7892/boris.54331

URI:

https://boris.unibe.ch/id/eprint/54331

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