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Comparison of Different Mobilization Strategies for Capillary Isoelectric Focusing of Ovalbumin Variants.
MedLine Citation:
PMID:  25358859     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
One pressure and three chemical mobilization strategies have been optimized and tested for two-step capillary isoelectric focusing with UV detection with simultaneous refining of the composition of carrier ampholytes as well as of anodic and cathodic spacers. The comparison of individual mobilization strategies was performed on basis of model proteins and peptides covering a pI range of 4.1-10.0 finally targeting an acidic major food allergen, i.e., ovalbumin. Resolution was improved by combining Pharmalyte 3-10 with Pharmalyte 5-6 with concentration adjustment of carrier ampholytes and the anodic and cathodic spacer, respectively. Analytes within pI 5-6 but not ovalbumin were prone to artificial peak duplication under selected capillary isoelectric focusing conditions due to retardation during focusing. l-Arginine and iminodiacetic acid were included as spacer to prevent drifts of the pH gradient and optionally block the distal capillary part. l-Arginine affected the baseline in the acidic regime in some instances by introducing irregularities that interfered with ovalbumin. Cathodic mobilization with an acidic zwitterion provided best selectivity for ovalbumin and was successfully applied for characterization of three commercial products of ovalbumin revealing differences between the respective profiles. Up to 12 different fractions situated between pI 4.51 and 4.72 could be addressed. This article is protected by copyright. All rights reserved.
Authors:
Theresa Kristl; Hanno Stutz
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-10-31
Journal Detail:
Title:  Journal of separation science     Volume:  -     ISSN:  1615-9314     ISO Abbreviation:  J Sep Sci     Publication Date:  2014 Oct 
Date Detail:
Created Date:  2014-10-31     Completed Date:  -     Revised Date:  2014-11-2    
Medline Journal Info:
Nlm Unique ID:  101088554     Medline TA:  J Sep Sci     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
This article is protected by copyright. All rights reserved.
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