Factors that influence hemoglobin (Hb)AIc synthesis by intact erythrocytes were studied in vitro. After incubation cells were lysed, and hemoglobins were separated by isoelectric focusing on polyacrylamide slab gels and quantitated by microdensitometry. HbAIc increased with time, glucose concentrations (5-500 mM), and incubation temperature (4°-37°C). Low temperatures allowed prolonged incubations with minimal hemolysis. At 4°C HbAIc increased linearly with time for 6 wk; after incubation at the highest glucose concentration, HbAIc comprised 50% of total hemoglobin.
Kenneth M. Spicer, Robert C. Allen, David Hallett, Maria G. Buse