Glavanovich M H, Carr P W
Department of Chemistry, University of Minnesota, Minneapolis 55455-0431.
Anal Chem. 1994 Aug 1;66(15):2584-9. doi: 10.1021/ac00087a025.
The goal of this work was to develop a generic approach for producing affinity chromatographic columns which can be regenerated. Concanavalin A (Con A) was immobilized adsorptively by an in situ method onto a zirconium dioxide (zirconia) chromatographic support and used to resolve chromophorically labeled monosaccharides. The Con A was then removed from the zirconia by flushing with base. The same column was regenerated by applying a fresh aliquot of Con A. This cycle was repeated several times to demonstrate consistency in the loading capacity and the stability of the underlying zirconia support. Finally we used glutaraldehyde to cross-link the Con A to increase the long-term stability of the column. Hydrolyzing the protein with acid allowed it to be removed under alkaline conditions and the column regenerated simply by adding more Con A followed by glutaraldehyde cross-linking.
这项工作的目标是开发一种生产可再生亲和色谱柱的通用方法。伴刀豆球蛋白A（Con A）通过原位法吸附固定在二氧化锆（氧化锆）色谱支持物上，并用于分离发色团标记的单糖。然后通过用碱冲洗从氧化锆中去除Con A。通过加入新鲜的Con A等分试样使同一色谱柱再生。重复这个循环几次，以证明负载能力的一致性和底层氧化锆支持物的稳定性。最后，我们使用戊二醛使Con A交联，以提高色谱柱的长期稳定性。用酸水解蛋白质使其在碱性条件下被去除，并且通过简单地添加更多Con A然后进行戊二醛交联来使色谱柱再生。
