HbA1c和HbA0的分离纯化

doi:10.3969/j.issn.1673-8640.2013.10.010

检验医学 ›› 2013, Vol. 28 ›› Issue (10): 904-907.DOI: 10.3969/j.issn.1673-8640.2013.10.010

• 技术研究与评价.论著 • 上一篇下一篇

HbA_1c和HbA₀的分离纯化

涂国华¹，王碧筠¹，朱云娇¹，秦瑛¹，姜旭淦²

1.无锡市滨湖区中医院检验科，江苏无锡 214121；2.江苏大学基础医学与医学技术学院，江苏镇江 212013

收稿日期:2012-11-15 出版日期:2013-10-30 发布日期:2013-10-29
通讯作者: 姜旭淦，联系电话:0511-85038483。
作者简介:涂国华，男，1978年生，硕士，主管技师，主要从事临床检验工作。

Separation and purification of HbA_1c and HbA₀

TU Guohua¹,WANG Bijun¹,ZHU Yunjiao¹,QIN Ying¹,JIANG Xugan².

1.Department of Clinical Laboratory,Binhu District Traditional Chinese Medical Hospital,Jiangsu Wuxi 214121,China；2. School of Medical Science and Laboratory Medicine,Jiangsu University,Jiangsu Zhenjiang 212013,China

Received:2012-11-15 Online:2013-10-30 Published:2013-10-29

摘要/Abstract

摘要：

目的从健康人红细胞分离并纯化血红蛋白A1c(HbA_1c)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HbA_1c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HbA_1c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，HbA_1c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HbA_1c和HbA₀。

关键词: 血红蛋白A_1c, 血红蛋白A₀, 离子交换树脂, 亲和层析

Abstract:

Objective To separate and purify hemoglobin A_1c(HbA_1c) 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. HbA_1c 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 HbA_1c and HbA₀ from healthy subjects′ 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 HbA_1c 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 HbA_1c and HbA₀.

Key words: Hemoglobin A_1c, Hemoglobin A₀, Ion exchange resin, Affinity chromatography

中图分类号:

R446.1

涂国华，王碧筠，朱云娇，秦瑛，姜旭淦. HbA_1c和HbA₀的分离纯化[J]. 检验医学, 2013, 28(10): 904-907.

TU Guohua,WANG Bijun,ZHU Yunjiao,QIN Ying,JIANG Xugan.. Separation and purification of HbA_1c and HbA₀[J]. , 2013, 28(10): 904-907.

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HbA_1c和HbA₀的分离纯化

Separation and purification of HbA_1c and HbA₀

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