Gradient Chromatofocusing. Versatile pH Gradient Separation of Proteins in Ion-Exchange HPLC:  Characterization Studies

2002 ◽  
Vol 74 (21) ◽  
pp. 5641-5649 ◽  
Author(s):  
Lian Shan ◽  
David J. Anderson
Keyword(s):  
Ion Exchange ◽  
Ph Gradient ◽  
Ion Exchange Hplc ◽  
Characterization Studies ◽  
Gradient Chromatofocusing ◽  
Separation Of Proteins ◽  
Gradient Separation

Isoelectric point separation of proteins by capillary pH-gradient ion-exchange chromatography

2004 ◽  
Vol 1025 (2) ◽  
pp. 217-226 ◽  
Author(s):  
Thomas Andersen ◽  
Milaim Pepaj ◽  
Roger Trones ◽  
Elsa Lundanes ◽  
Tyge Greibrokk
Keyword(s):  
Ion Exchange ◽  
Isoelectric Point ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Ph Gradient ◽  
Point Separation ◽  
Separation Of Proteins

Selection of pH-related parameters in ion-exchange chromatography using pH-gradient operations

2008 ◽  
Vol 1194 (1) ◽  
pp. 22-29 ◽  
Author(s):  
Tangir Ahamed ◽  
Sreekanth Chilamkurthi ◽  
Beckley K. Nfor ◽  
Peter D.E.M. Verhaert ◽  
Gijs W.K. van Dedem ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Ph Gradient ◽  
Selection Of

Mixed-Bed Ion Exchange Chromatography Employing a Salt-Free pH Gradient for Improved Sensitivity and Compatibility in MudPIT

2013 ◽  
Vol 85 (14) ◽  
pp. 6608-6616 ◽  
Author(s):  
Geert P. M. Mommen ◽  
Hugo D. Meiring ◽  
Albert J. R. Heck ◽  
Ad P. J. M. de Jong
Keyword(s):  
Ion Exchange ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Ph Gradient ◽  
Mixed Bed Ion Exchange

Affinity Binding Assay of Glycohemoglobin by Two-Dimensional Centrifugation Referenced to Hemoglobin A1C

1992 ◽  
Vol 38 (12) ◽  
pp. 2372-2379 ◽  
Author(s):  
M Fiechtner ◽  
J Ramp ◽  
B England ◽  
M A Knudson ◽  
R R Little ◽  
...  
Keyword(s):  
Ion Exchange ◽  
Hemoglobin A1c ◽  
Solid Phase ◽  
Capillary Tube ◽  
Glass Capillary ◽  
Agarose Beads ◽  
Automated Assay ◽  
Regression Parameters ◽  
Ion Exchange Hplc ◽  
Blood Specimens

Abstract We describe an automated assay of glycohemoglobin performed with the Abbott Vision analyzer. The assay is based on batch affinity-extraction with 3-aminophenylboronic acid-derivatized agarose beads. Reagents are packaged in a disposable test pack. Whole-blood specimens are hemolyzed with saponin within a glass capillary tube inserted into the test pack. The sample is automatically diluted with, mixed with, and separated from the solid-phase reagent. Bichromatic absorbance readings are used to calculate the percentage of hemoglobin bound. Based on the linear correlation between affinity-measured glycohemoglobin and HPLC-measured hemoglobin A1c, the percentage of hemoglobin bound is converted to a "standardized %HbA1c" result by use of regression parameters stored during a calibration run. The combination of affinity methodology with standardization by reference to HPLC produces values directly comparable with those obtained by methods specific for HbA1c. The method produces 10 results within 15 min. The assay operates with CVs < 5%, and the results correlate highly with those by ion-exchange and affinity minicolumn methods, and by ion-exchange HPLC.

scholarly journals Calibration of ion-exchange HPLC measurements of glycohemoglobin: effect on interassay precision

1997 ◽  
Vol 43 (12) ◽  
pp. 2353-2357 ◽  
Author(s):  
Erik Gerlo ◽  
Frans Gorus
Keyword(s):  
Ion Exchange ◽  
Interlaboratory Comparisons ◽  
Conversion Factors ◽  
Hplc System ◽  
Ion Exchange Hplc ◽  
Assay Precision ◽  
Interassay Precision ◽  
Control Samples

Abstract To investigate the effect of calibration with lyophilized calibrators on the interassay precision of glycohemoglobin (glyHb) measurements, we used an ion-exchange HPLC system equipped with a Pharmacia Mono S® HR 5/5 column. Calibration of analytical runs substantially increased interassay variation (CV), from 1.7% to 4.4% and from 0.9% to 3.2% for control samples with low (6.5%) and high (14%) glyHb percentages, respectively. Standardization of glyHb results, though essential for interlaboratory comparisons, should not be done at the expense of assay precision, as may occur with thoughtless use of lyophilized calibrators. We therefore recommend the use of carefully determined conversion factors for standardization of glyHb results obtained with ion-exchange HPLC systems that are capable of excellent long-term interassay precision.

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