Weiss A J, McElhinney S A, Blankstein L A
Millipore Corporation, Bedford, MA 01730.
Biotechniques. 1989 Oct;7(9):1012-6.
With the model ligands studies, which included IgG, HSA, streptavidin, MEA and amine-modified DNA, it was possible to enhance the rate of covalent immobilization by using nucleophilic acylation reaction catalysts. Imidazole, triazole and 2-hydroxypyridine are readily available catalysts that are effective when immobilizing immunoglobins. 4-N,N,Dimethylaminopyridine (DMAP) as a co-reactant or as a prereactant is a potent rate enhancer with all of the molecules that were examined. The precise protocol to be used is probably best derived empirically. In addition to optimizing the amount of ligand bound or the amount of time necessary to bind a fixed quantity of ligand, it is likely that the retained functionality of the ligand may be affected by the use of reaction catalysts.
通过对包括IgG、HSA、链霉亲和素、MEA和胺修饰DNA在内的模型配体进行研究,利用亲核酰化反应催化剂可以提高共价固定的速率。咪唑、三唑和2-羟基吡啶是易于获得的催化剂,在固定免疫球蛋白时很有效。4-N,N-二甲基氨基吡啶(DMAP)作为共反应物或预反应物,对所有检测的分子都是一种有效的速率增强剂。具体使用的方案可能最好通过经验得出。除了优化结合的配体量或结合固定量配体所需的时间外,配体的保留功能可能会受到反应催化剂使用的影响。