Separation of glycosylated haemoglobins using immobilized phenylboronic acid. Effect of ligand concentration, column operating conditions, and comparison with ion-exchange and isoelectric-focusing
Autor: | F A Middle, Peter D.G. Dean, A Bannister, A J Bellingham |
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Rok vydání: | 1983 |
Předmět: |
Magnesium Chloride
chemistry.chemical_element Buffers Ligands Biochemistry Chromatography Affinity Haemoglobin A chemistry.chemical_compound Affinity chromatography Diabetes Mellitus Humans Magnesium Phenylboronic acid Molecular Biology Glycated Hemoglobin Chromatography Ion exchange Chemistry Isoelectric focusing Haemoglobin A1c Ligand Temperature Cell Biology Hydrogen-Ion Concentration Chromatography Ion Exchange Boronic Acids Isoelectric Focusing Research Article |
Zdroj: | Biochemical Journal. 209:771-779 |
ISSN: | 0264-6021 |
DOI: | 10.1042/bj2090771 |
Popis: | Haemoglobins from diabetic and non-diabetic individuals have been separated by affinity chromatography using immobilized phenylboronate, which interacts specifically with diol-containing compounds such as glycosylated haemoglobin. The effects of ligand concentration, flow rate, column geometry, preincubation of sample, buffer composition and temperature have been investigated. Significant correlation was found between results from affinity-chromatography and ion-exchange and isoelectric-focusing methods. Isoelectric-focusing of the haemoglobin fractions obtained from affinity chromatography indicate that, in addition to haemoglobin A1c, some haemoglobin A is also bound to immobilized phenylboronic acid. Assays of haemolysates obtained from red blood cells incubated in glucose solutions suggest that unstable pre-(haemoglobin A1c) does not interfere. The assay is not affected by the presence of haemoglobin F. |
Databáze: | OpenAIRE |
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