Hemoglobin Raleigh as the cause of a falsely increased hemoglobin A(1C) in an automated ion-exchange HPLC method

Dan Chen, Dan L. Crimmins, Fong Fu Hsu, Frederik P. Lindberg, Mitchell G. Scott

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

Irreversible glycation of the hemoglobin A0 (HbA0) β chain leads to the production of HbA(1C) which can be used to monitor long-term blood glucose control in patients with diabetes mellitus. HbA(1C) is less positively charged than nonglycated HbA0, and this decrease in charge is the basis of ion-exchange and electrophoretic methods that measure HbA(1C). We recently identified a sample that appeared to contain 46% HbA(1C) by an automated ion-exchange HPLC method (Bio-Rad Variant(TM)) but only 3.8% by an immunoinhibition latex agglutination method. A combination of traditional and mass spectrometric protein analysis and genomic DNA analysis of the Hb β chain and genes revealed that the patient was heterozygotic for Hb-Raleigh, a variant containing a valine → alanine substitution at position 1 of the β chain. The amino-terminal alanine in this variant Hb is posttranslationally modified by acetylation, leading to a charge difference similar to glycation and making the behavior of HbA(1C) and Hb Raleigh virtually identical in the ion-exchange HPLC method. This observation suggests that it is important to confirm HbA(1C) values in excess of 15% especially if they are not consistent with the clinical picture, by an independent HbA(1C) method such as immunoassay or boronic acid affinity chromatography. However, for this particular variant Hb, even these latter-methods might be misleading, because the acetylated N-terminal amino acid of the Hb-Raleigh β chain cannot be glycated.

Original languageEnglish
Pages (from-to)1296-1301
Number of pages6
JournalClinical chemistry
Volume44
Issue number6
StatePublished - Jun 30 1998

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