This feasibility study deals with the use of preparative capillary isotachophoresis (CITP), operating in a discontinuous fractionation mode, to the separations and isolations of glycoforms of recombinant human erythropoietin (rhEPO). The preparative CITP separations were monitored by capillary zone electrophoresis (CZE) with a hydrodynamically closed separation unit. Such a CZE system, suppressing fluctuations of the migration data linked with fluctuations of EOF and hydrodynamic flow, made possible to evaluate and compare the preparative CITP separations performed within a longer time frame. Preparative CITP, carried out in the separation unit with coupled columns of enhanced sample loadability, separating 100 microg of rhEPO in a run lasting ca. 30 min, gave the production rate higher than 55 ng/s for the rhEPO glycoforms. The preparative separations included valve isolations of the glycoforms from the ITP stack into four or six fractions. Such numbers of the fractions corresponded to typical numbers of the major glycoform peaks as resolved in CZE of rhEPO. With respect to close effective mobilities of the glycoforms and a multicomponent nature of rhEPO, the fractions contained mixtures of glycoforms with the dominant glycoforms enriched 10-100-fold, relative to the original rhEPO sample.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1002/elps.200500044 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
The minimum information required for a glycomics experiment (MIRAGE): reporting guidelines for capillary electrophoresis.
Glycobiology
June 2022
Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston University Medical Campus, 715 Albany Street, Boston, MA 02118, United States.
The Minimum Information Required for a Glycomics Experiment (MIRAGE) is an initiative to standardize the reporting of glycoanalytical methods and to assess their reproducibility. To date, the MIRAGE Commission has published several reporting guidelines that describe what information should be provided for sample preparation methods, mass spectrometry methods, liquid chromatography analysis, exoglycosidase digestions, glycan microarray methods, and nuclear magnetic resonance methods. Here, we present the first version of reporting guidelines for glyco(proteo)mics analysis by capillary electrophoresis (CE) for standardized and high-quality reporting of experimental conditions in the scientific literature.
View Article and Find Full Text PDFPolarization Control via He-Ion Beam Induced Nanofabrication in Layered Ferroelectric Semiconductors.
ACS Appl Mater Interfaces
March 2016
The Institute for Functional Imaging of Materials and the Center for Nanophase Materials Sciences, ‡Center for Nanophase Materials Sciences, and ⊥Materials Sciences and Technology Division, Oak Ridge National Laboratory , Oak Ridge, Tennessee 37831, United States.
Rapid advances in nanoscience rely on continuous improvements of material manipulation at near-atomic scales. Currently, the workhorse of nanofabrication is resist-based lithography and its various derivatives. However, the use of local electron, ion, and physical probe methods is expanding, driven largely by the need for fabrication without the multistep preparation processes that can result in contamination from resists and solvents.
View Article and Find Full Text PDFUnlabelled: Study objective was to evaluate and to compare glutamine, glutamate and citrulline plasma levels (Glu-p, Gl-p, Cit-p) with electrical activity indicators (EA) from the proximal small intestine in patients in critical condition (CC).
Materials And Methods: 27 patients in CC (15 women, 12 men, 70 +/- 14 years, from them "therapeutic profile" patients--12, "surgical"--15) have been surveyed and treated. At admission to reanimatology department (RD) Glu-p, Gl-p, Cit-p (by highly effective liquid chromatography), relative duodenal and jejunum EA--by peripheral electrogastrography--have been measured.
This paper deals with an analytical method based on two dimensional capillary electrophoresis (CITP-CZE coupling) with simple UV-detection for the determination of a highly effective drug - varenicline. The method was elaborated with the possibility of its future connection with an advanced detection ending - mass spectrometry. The electrolytes for the CITP (LE = 10 mM NH4Ac + 5 mM HAc + 0,05% m-HEC; TE = 10 mM HAc) and CZE (BGE = 20 mM HAc) separation of varenicline were selected considering such requirements.
View Article and Find Full Text PDFModest elevation in BNP in asymptomatic hypertensive patients reflects sub-clinical cardiac remodeling, inflammation and extracellular matrix changes.
PLoS One
May 2013
Heart Failure Unit, St Vincent's University Hospital, Elm Park, Dublin, Ireland.
In asymptomatic subjects B-type natriuretic peptide (BNP) is associated with adverse cardiovascular outcomes even at levels well below contemporary thresholds used for the diagnosis of heart failure. The mechanisms behind these observations are unclear. We examined the hypothesis that in an asymptomatic hypertensive population BNP would be associated with sub-clinical evidence of cardiac remodeling, inflammation and extracellular matrix (ECM) alterations.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!