Siemiatkoski Joseph, Lyubarskaya Yelena, Houde Damian, Tep Samnang, Mhatre Rohin
Analytical Development, Biogen Idec, Inc, Cambridge, MA 02142, USA.
Carbohydr Res. 2006 Feb 27;341(3):410-9. doi: 10.1016/j.carres.2005.11.024. Epub 2005 Dec 27.
A comparison of three techniques for quantitative analysis of galactosylation present on immunoglobulins is described. ESIMS, MALDI-TOF MS, and anion-exchange chromatography with fluorescence detection were evaluated in terms of repeatability, limit of quantitation, selectivity, and linearity. A recombinant monoclonal IgG was enzymatically modified in vitro to produce essentially completely galactosylated and degalactosylated forms of the immunoglobulin. Samples of known galactosylation levels were prepared by mixing the modified forms with the native form of the immunoglobulin. Good repeatability and linearity were demonstrated for all three assays (RSDs<1.0%, correlation coefficients>0.99). Differences in selectivity, sensitivity, and other performance aspects of the three techniques are also discussed in this paper.
本文描述了三种用于定量分析免疫球蛋白上半乳糖基化的技术的比较。从重复性、定量限、选择性和线性方面对电喷雾离子化质谱（ESIMS）、基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）和带荧光检测的阴离子交换色谱法进行了评估。一种重组单克隆IgG在体外经酶修饰，以产生基本上完全半乳糖基化和去半乳糖基化形式的免疫球蛋白。通过将修饰形式与免疫球蛋白的天然形式混合制备已知半乳糖基化水平的样品。所有三种测定方法均显示出良好的重复性和线性（相对标准偏差<1.0%，相关系数>0.99）。本文还讨论了这三种技术在选择性、灵敏度和其他性能方面的差异。
