Kobold U, Jeppsson J O, Dülffer T, Finke A, Hoelzel W, Miedema K
Boehringer Mannheim GmbH Lab Diagnostics, Research Center Tutzing, Germany.
Clin Chem. 1997 Oct;43(10):1944-51.
A reference method that specifically measures hemoglobin (Hb) A1c is an essential part of the reference system for the international standardization of Hb A1c/glycohemoglobin. We have developed a new method for quantification, based on the specific N-terminal residue of the hemoglobin beta-chains. Enzymatic cleavage of the intact hemoglobin molecule with endoproteinase Glu-C has been optimized to obtain the beta-N-terminal hexapeptides of Hb A1c and Hb A0. These peptides have been separated by reversed-phase HPLC and quantitated by electrospray ionization-mass spectrometry (method A) or by capillary electrophoresis (method B). With these peptides and hyphenated separation techniques, it has been possible to overcome the insufficient resolution of currently used protein separation systems for Hb A1c.
一种专门测量糖化血红蛋白（Hb）A1c的参考方法是Hb A1c/糖化血红蛋白国际标准化参考系统的重要组成部分。我们基于血红蛋白β链的特定N端残基开发了一种新的定量方法。用内肽酶Glu-C对完整血红蛋白分子进行酶切已得到优化，以获得Hb A1c和Hb A0的β-N端六肽。这些肽已通过反相高效液相色谱法分离，并通过电喷雾电离质谱法（方法A）或毛细管电泳法（方法B）进行定量。利用这些肽和联用分离技术，得以克服目前用于Hb A1c的蛋白质分离系统分辨率不足的问题。
