DEVELOPMENT OF DIHYDRAZIDE-ACTIVATED SILICA SUPPORTS FOR HIGHPERFORMANCE AFFINITY-CHROMATOGRAPHY
DEVELOPMENT OF DIHYDRAZIDE-ACTIVATED SILICA SUPPORTS FOR HIGHPERFORMANCE AFFINITY-CHROMATOGRAPHY
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DOI：
10.1016/0021-9673(94)80332-3
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发表时间：
1994-05-27
影响因子：
4.1
通讯作者：
HAGE, DS
中科院分区：
文献类型：
--
作者：
RUHN, PF;GARVER, S;HAGE, DS
建立了一种用于高效亲和层析(HPAC)的二肼基活性硅胶的制备方法。这种载体是通过氧化二元醇键合的二氧化硅并与草酸或己二肼反应制成的。对合成过程中涉及的步骤进行了研究，并用FTIR进行了确认。在优化载体制备时考虑的项目包括二肼的加入量和反应时间或使用的pH。通过改变二醇键合二氧化硅的氧化程度来控制二肼基双官能团附着。该载体已成功应用于氧化抗体、辣根过氧化物酶和转移核糖核酸的固定化。在每种情况下，数据都表明固定化是通过特定部位的偶联，而不是非特定的吸附。二肼活化的二氧化硅在制备后在5-25℃下保存2-6周后被发现是稳定的。在HPAC中常用的各种溶剂的存在下，氧化生物分子与该载体之间的连接至少稳定了一个月。
A method of preparing dihydrazide-activated silica was developed for use in high-performance affinity chromatography (HPAC). This support was made by oxidizing diol-bonded silica and reacting it with oxalic or adipic dihydrazide. The steps involved in this synthesis were studied and confirmed by FTIR. Items considered in optimizing the preparation of the support included the amount of dihydrazide added and the reaction time or pH used. Control of dihydrazide bifunctional attachment was obtained by varying the extent of diol-bonded silica oxidation. This support was successfully used in the immobilization of oxidized antibodies, horse radish peroxidase (a glycoenzyme) and transfer RNA. In each case, data indicated that immobilization was through site specific coupling rather than non-specific adsorption. Dihydrazide-activated silica was found to be stable for 2-6 weeks after preparation when stored at 5 to 25 degrees C. The linkage between oxidized biomolecules and this support was stable for at least one month in the presence of various solvents commonly used in HPAC.