HbA1c和HbA0的分离纯化

目的从健康人红细胞分离并纯化血红蛋白A1c（HbA1c）和血红蛋白A0（HbA0）。方法采集健康人全血，分离红细胞并制备溶血液，采用弱酸阳离子交换层析法分离HbA1c和HbA0，并用硼酸琼脂糖亲和层析法分别纯化。结果健康人红细胞溶血液采用弱酸阳离子交换层析法手工装柱分离提取HbA1c和HbA0峰值部分，高效液相色谱（HPLC）法测定纯度分别为79．78％和97．42％；再经硼酸琼脂糖亲和层析分别纯化，HbA1c纯度可达94．3％，HbA0纯度可达99．7％。结论单一的离子交换方法分离糖化Hb并不理想，必须与亲和层析结合，才能得到较高纯度HbA1c和HbA0。

Separation and purification of HbA1c and HbA0

Objective To separate and purify hemoglobin A2c （ HbA1c ） and hemoglobin A0 （ HbA0 ） from red blood cells of healthy subjects. Methods Collecting whole blood from healthy subjects, the red blood cells were separated and prepared for hemolysis. HbA1c and HbA0 were separated by weak acid positive ion exchange chromatography and purified by boric acid agarose affinity chromatography respectively. Results Hand-loaded column with weak acid positive ion exchange chromatography was used to separate and collect the peaks of HbA1c and HbAo from healthy sub- jects＇ red blood cell hemolysis, and the purities were 79.78% and 97.42% by high performance liquid chromatography （HPLC） respectively. After being purified by boric acid agarose affinity chromatography,the final purities of HbA1c and HbA0 were 94.3% and 99.7% respectively. Conclusions The single ion exchange chromatography for separating the glycosylated hemoglobin is not ideal, and it must be combined with affinity chromatography to obtain a high purities of HbA1c and HbA0.