National Research Council of Italy, Istituto di Scienze e Tecnologie Chimiche (SCITEC-CNR), Milan, Italy.
Dipartimento di Scienze Farmaceutiche (DISFARM), Università degli Studi di Milano, Milan, Italy.
Methods Mol Biol. 2023;2578:27-39. doi: 10.1007/978-1-0716-2732-7_3.
The analytical performance of the microarray technique in screening the affinity and reactivity of molecules toward a specific target is highly affected by the coupling chemistry adopted to bind probes to the surface. However, the surface functionality limits the biomolecules that can be attached to the surface to a single type of molecule, thus forcing the execution of separate analyses to compare the performance of different species in recognizing their targets. Here, we introduce a new N,N-dimethylacrylamide-based polymeric coating, bearing simultaneously different functionalities (N-acryloyloxysuccinimide and azide groups) to allow an easy and straightforward method to co-immobilize proteins and oriented peptides on the same substrate. The bifunctional copolymer has been obtained by partial post-polymerization modification of the functional groups of a common precursor. This strategy represents a convenient method to reduce the number of analyses, therefore possible systematic or random errors, besides offering a drastic shortage in time, reagents, and costs.
微阵列技术在筛选分子对特定靶标的亲和力和反应性方面的分析性能,高度受到用于将探针结合到表面的偶联化学的影响。然而,表面功能限制了可以附着到表面的生物分子的类型为单一类型的分子,从而迫使执行单独的分析以比较不同物种识别其靶标的性能。在这里,我们引入了一种新的基于 N,N-二甲基丙烯酰胺的聚合物涂层,其同时具有不同的功能(丙烯酰氧基琥珀酰亚胺和叠氮基团),以允许一种简单而直接的方法将蛋白质和定向肽共同固定在相同的基质上。双功能共聚物是通过对常见前体的功能基团进行部分后聚合修饰而获得的。这种策略代表了一种减少分析数量的简便方法,因此可能会减少系统或随机误差,同时大大缩短时间、试剂和成本。