López-Gallego Fernando, Guisán Jose M, Betancor Lorena
Institute of Catalysis, CSIC, CAMPUS UAM-Cantoblanco, Madrid, Spain.
Methods Mol Biol. 2013;1051:33-41. doi: 10.1007/978-1-62703-550-7_3.
In this chapter, we describe different approaches for the utilization of glutaraldehyde in protein immobilization. First, we focus on the covalent attachment of proteins to glutaraldehyde-activated matrixes. We describe conditions for the synthesis of such supports and provide an example of the immobilization and stabilization of fructosyltransferase. We also describe how glutaraldehyde may be used for the cross-linking of protein-protein aggregates and protein adsorbed onto amino-activated matrixes. In these cases, glutaraldehyde bridges either two lysine groups from different proteic molecules or a lysine from the protein structure and an amine group from the support. Examples of cross-linking are given for the immobilization of DAAO on different amino-activated supports.
在本章中,我们描述了戊二醛在蛋白质固定化中的不同应用方法。首先,我们着重于蛋白质与戊二醛活化基质的共价连接。我们描述了此类载体的合成条件,并给出了果糖基转移酶固定化和稳定化的示例。我们还描述了戊二醛如何用于蛋白质 - 蛋白质聚集体以及吸附在氨基活化基质上的蛋白质的交联。在这些情况下,戊二醛桥接不同蛋白质分子的两个赖氨酸基团,或者蛋白质结构中的赖氨酸和载体上的胺基。给出了在不同氨基活化载体上固定化DAAO的交联示例。