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PMID: 6870791 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Separation of glycosylated haemoglobins using immobilized phenylboronic acid. Effect of ligand concentration, column operating conditions, and comparison with ion-exchange and isoelectric-focusing.

The Biochemical journal ·Vol. 209 ·No. 3 ·1983-03-01 ·Pages 771-9

Middle FA, Bannister A, Bellingham AJ, Dean PD

Abstract

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.

MeSH Terms
Boronic Acids Buffers Chromatography, Affinity/methods Chromatography, Ion Exchange Diabetes Mellitus/blood Glycated Hemoglobin A/isolation & purification Humans Hydrogen-Ion Concentration Isoelectric Focusing Ligands Magnesium Magnesium Chloride Temperature
Chemicals
Boronic Acids Buffers Glycated Hemoglobin A Ligands Magnesium Chloride Magnesium benzeneboronic acid
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Middle F A
Bannister A
Bellingham A J
Dean P D
References (19)
19 references, click to expand
  1. Synthesis of cellulose derivatives containing the dihydroxyboryl group and a study of their capacity to form specific complexes with sugars and nucleic acid components.
    Biochemistry. 1970 Oct 27;9(22):4396-401 PMID: 5472711
  2. Hemoglobin components in patients with diabetes mellitus.
    N Engl J Med. 1971 Feb 18;284(7):353-7 PMID: 5539916
  3. Studies on the interactions of nucleotides, polynucleotides, and nucleic acids with dihydroxyboryl-substituted celluloses.
    Biochemistry. 1972 Sep 12;11(19):3623-8 PMID: 5053763
  4. An improved method for the purification of tRNA by chromatography on dihydroxyboryl substituted cellulose.
    Nucleic Acids Res. 1975 Jun;2(6):853-64 PMID: 1096084
  5. In vitro synthesis of hemoglobin AIc.
    FEBS Lett. 1976 Dec 1;71(2):356-60 PMID: 12012
  6. Glycosylated minor components of human adult hemoglobin. Purification, identification, and partial structural analysis.
    J Biol Chem. 1978 Apr 10;253(7):2327-32 PMID: 632272
  7. Glycosylated hemoglobins: increased glycosylation of hemoglobin A in diabetic patients.
    Diabetes. 1979 Apr;28(4):337-40 PMID: 437373
  8. Structural heterogeneity of human hemoglobin A due to nonenzymatic glycosylation.
    J Biol Chem. 1979 May 25;254(10):3892-8 PMID: 438166
  9. Sites of nonenzymatic glycosylation of human hemoglobin A.
    J Biol Chem. 1980 Apr 10;255(7):3120-7 PMID: 7358733
  10. Increased glycosylation of serum albumin in diabetes mellitus.
    Diabetes. 1980 Jun;29(6):417-22 PMID: 6991333
  11. Rapid changes in chromatographically determined haemoglobin A1c induced by short-term changes in glucose concentration.
    Diabetologia. 1980 Aug;19(2):130-6 PMID: 7418966
  12. Sources of variation in the column-chromatographic determination of glycohemoglobin (HbA1).
    Clin Chem. 1981 Jan;27(1):94-9 PMID: 7449129
  13. Affinity chromatography of membrane proteins on dihydroxyboryl-matrix gel.
    Biochem Soc Trans. 1981 Feb;9(1):137-9 PMID: 7215645
  14. Kinetic analysis of the nonenzymatic glycosylation of hemoglobin.
    J Biol Chem. 1981 May 25;256(10):5204-8 PMID: 7228877
  15. The importance of determining irreversibly glycosylated hemoglobin in diabetics.
    Diabetes. 1981 Jul;30(7):607-12 PMID: 7250531
  16. Evaluation of glycosylated hemoglobin diabetic patients.
    Diabetes. 1981 Jul;30(7):613-7 PMID: 7250532
  17. The carbohydrate moiety in hemoglobin A1C is present in the ring form.
    FEBS Lett. 1981 Nov 30;135(1):145-7 PMID: 7319032
  18. Fractionation of membrane proteins on phenylboronic acid-agarose.
    Biochem J. 1982 Jul 1;205(1):167-71 PMID: 6181777
  19. The measurement of glycosylated hemoglobin in man and animals by aminophenylboronic acid affinity chromatography.
    Diabetes. 1982 Aug;31(8 Pt 1):701-5 PMID: 7160542
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2090771
Published
1983-03-01
Pages
771-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1154156
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2090771 PMC Full Text View journal details
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