Affinity Chromatography: A Precise Method for Glycosylated Albumin Estimation

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Abstract
A method is described for the estimation, in plasma, of glycosylated albumin, using the commercially available affinity chromatographic material Glycogel B (immobilised m-aminophenylboronic acid on an agarose support) to separate the glycosylated and non-glycosylated albumin, followed by the analysis of albumin concentration using rocket Immunoelectrophoresis. Glycosylated albumin and glycosylated haemoglobin showed good correlation (r=0·91) when both are estimated by affinity chromatography. The glycosylated albumin method displays good within-batch (2·8–4·4% CV) and between-batch (2·9–5·4% CV) precision, and the method is not affected by haemolysis. Using this method a reference range of 2·0–5·4% was found for glycosylated albumin. Levels of glycosylated albumin in diabetics (10·06±3·23%) were found to be significantly higher ( P<0·001) than those in non-diabetics (3·72±0·85%). It was found that loading more than 3 mg of albumin on to a 1 mL affinity column must be avoided, as this appears to overload the column.