Entrapment of Proteins Within Columns for High-Performance Affinity Chromatography.
Entrapment of Proteins Within Columns for High-Performance Affinity Chromatography.
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高效亲和色谱柱内蛋白质的包埋。
DOI：
10.1007/978-1-0716-2176-9_14
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Hage,DavidS
中科院分区：
文献类型：
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作者：
Poddar,Saumen;Sharmeen,Sadia;Hage,DavidS
包埋是一种非共价固定方法，其能够将大的生物结合剂（例如蛋白质）置于支持物内而不改变结合剂的结构。本章描述了一种柱上包埋方法，可用于蛋白质和HPLC级硅胶制备高效液相色谱柱。在该方法中，通过使用氧化糖原作为封端剂，蛋白质被捕获在二酰肼活化的二氧化硅载体内。该方法允许蛋白质以可溶形式放置在支持物内，并且几乎没有或没有活性损失。本章描述了该方法所需的方法和试剂，沿着了一些使用柱上蛋白包埋的色谱柱的应用。
Entrapment is a noncovalent immobilization method that enables a large biological binding agent, such as a protein, to be put within a support without modifying the structure of the binding agent. This chapter describes an on-column entrapment method that can be used with proteins and HPLC-grade silica to prepare columns for high-performance liquid chromatography. In this method, a protein is trapped within a dihydrazide-activated silica support by using oxidized glycogen as a capping agent. This method allows the protein to be placed within the support in a soluble form and with little or no loss of activity. The approach and reagents needed for this method are described in this chapter, along with some applications reported for columns that have been made using on-column protein entrapment.