Choe J, Zhang F, Wolff M W, Murhammer D W, Linhardt R J, Dordick J S
Department of Chemical Engineering, Rensselaer Polytechnic Institute, Troy, New York 12180, USA.
Biotechnol Bioeng. 2000 Dec 5;70(5):484-90.
A new method for the preparation of the glycoforms of bovine alpha(1)-acid glycoprotein (AGP) is described relying on affinity-reversed micellar extraction and separation (ARMES). This method has proven effective in separating structurally similar glycoproteins and separating glycoproteins from nonglycosylated proteins from natural sources. In this method, individual glycoforms complex with the lectin, concanavalin A (ConA) are extracted into an organic-phase reversed micellar solution formed by Aerosol OT (AOT). The purity of three AGP glycoforms isolated was assessed by hydroxyapatite high-performance liquid chromatography (HPLC), gel-permeation chromatography and SDS-PAGE. The glycan structure of the pure glycoforms was analyzed. Oligosaccharide mapping using capillary electrophoresis (CE) and PAGE showed the glycans obtained from each glycoform to be distinctly different. ARMES can be used for the semi-preparative scale resolution of the glycoforms of bovine AGP or other therapeutic glycoproteins.
本文描述了一种基于亲和反相胶束萃取与分离（ARMES）制备牛α(1)-酸性糖蛋白（AGP）糖型的新方法。该方法已被证明在分离结构相似的糖蛋白以及从天然来源的非糖基化蛋白中分离糖蛋白方面是有效的。在这种方法中，与凝集素伴刀豆球蛋白A（ConA）结合的各个糖型被萃取到由气溶胶OT（AOT）形成的有机相反相胶束溶液中。通过羟基磷灰石高效液相色谱（HPLC）、凝胶渗透色谱和SDS-PAGE对分离得到的三种AGP糖型的纯度进行了评估。对纯糖型的聚糖结构进行了分析。使用毛细管电泳（CE）和PAGE进行的寡糖图谱分析表明，从每种糖型获得的聚糖明显不同。ARMES可用于牛AGP或其他治疗性糖蛋白糖型的半制备规模分离。
