| Parameters affecting the separation of intact proteins in gradient-elution reversed-phase chromatography using poly(styrene-co-divinylbenzene) monolithic capillary columns. | |
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MedLine Citation:
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PMID: 20347095 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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An experimental study was performed to investigate the effects of column parameters and gradient conditions on the separation of intact proteins using styrene-based monolithic columns. The effect of flow rate on peak width was investigated at constant gradient steepness by normalizing the gradient time for the column hold-up time. When operating the column at a temperature of 60 degrees C a small C-term effect was observed in a flow rate range of 1-4 microL/min. However, the C-term effect on peak width is not as strong as the decrease in peak width due to increasing flow rate. The peak capacity increased according to the square root of the column length. Decreasing the macropore size of the polymer monolith while maintaining the column length constant, resulted in an increase in peak capacity. A trade-off between peak capacity and total analysis time was made for 50, 100, and 250 mm long monolithic columns and a microparticulate column packed with 5 microm porous silica particles while operating at a flow rate of 2 microL/min. The peak capacity per unit time of the 50mm long monolithic column with small pore size was superior when the total analysis time is below 120 min, yielding a maximum peak capacity of 380. For more demanding separations the 250 mm long monolith provided the highest peak capacity in the shortest possible time frame. |
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Authors:
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Frederik Detobel; Ken Broeckhoven; Joke Wellens; Bert Wouters; Remco Swart; Mario Ursem; Gert Desmet; Sebastiaan Eeltink |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-03-06 |
Journal Detail:
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Title: Journal of chromatography. A Volume: 1217 ISSN: 1873-3778 ISO Abbreviation: J Chromatogr A Publication Date: 2010 Apr |
Date Detail:
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Created Date: 2010-04-12 Completed Date: 2010-06-28 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9318488 Medline TA: J Chromatogr A Country: Netherlands |
Other Details:
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Languages: eng Pagination: 3085-90 Citation Subset: IM |
Copyright Information:
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2010 Elsevier B.V. All rights reserved. |
Affiliation:
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Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, 1050 Brussels, Belgium. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cattle Chickens Chromatography, High Pressure Liquid / methods* Chromatography, Reverse-Phase / methods* Horses Polystyrenes / chemistry* Proteins / isolation & purification* Proteomics / methods* |
| Chemical | |
Reg. No./Substance:
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0/Polystyrenes; 0/Proteins; 0/divinylbenzene-polystyrene copolymer |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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