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Determination of glycated apolipoprotein B in serum by a combination of affinity chromatography and
M Panteghini1, R Bonora, F Pagani
11st laboratory of Clinical Chemistry, Section of Clinical Chemistry and Enzymology, Spedali Civili, Brescia, Italy.
Abstract:
We have developed and optimized a procedure for the quantitation of non-enzymatically glycated apolipoprotein B (apo B). Glycated and non-glycated apo B were separated from serum using m-aminophenylboronate affinity chromatography, determined by immunophelometry and the percentage of glycated apo B was calculated. The measuring range of the assay was 2.9-185 mg/dL apo B. The within- and between-run coefficients of variation were < 7.4% and 14.6%, respectively, and recovery was > 98%. Free glucose in serum did not affect the results at concentrations below 25 mmol/L. In 45 non-diabetic subjects the mean concentration of glycated apo B was 4.3% (SD 1%). In type 1 (n = 17) and Type 2 (n = 60) diabetic patients the mean glycated apo B concentrations were 5.3% (SD 0.7%) and 5.9% (SD 1.1%), respectively, significantly higher than in controls (P < 0.001).