Zammatteo N, Girardeaux C, Delforge D, Pireaux J J, Remacle J
Laboratoire de Biochimie Cellulaire, Facultés Universitaires Notre-Dame de la Paix, rue de Bruxelles 61, Namur, 5000, Belgium.
Anal Biochem. 1996 Apr 5;236(1):85-94. doi: 10.1006/abio.1996.0135.
Amine functionalization of polystyrene microwells for covalent binding of DNA is described. Polystyrene support was first carboxylated by permanganate oxidation in diluted sulfuric acid. These functions were activated with water-soluble carbodiimide and grafted with N-methyl-1,3-propane diamine to introduce a free secondary amino group on the support. The samples were characterized by X-ray photoelectron spectroscopy and radiochemical assay. The conditions for covalent linkage of secondary amine on polystyrene microwells were optimized. Functionalized supports were used for covalent binding of a DNA capture probe for the detection of human cytomegalovirus in a sandwich hybridization assay. Sensitivity of the assay compared very well with a commercially available surface, Covalink-NH, microwell plate obtained by electromagnetic irradiation.
描述了用于DNA共价结合的聚苯乙烯微孔的胺功能化。聚苯乙烯载体首先在稀硫酸中通过高锰酸钾氧化进行羧基化。这些官能团用水溶性碳二亚胺活化，并与N-甲基-1,3-丙二胺接枝，以在载体上引入游离仲氨基。通过X射线光电子能谱和放射化学分析对样品进行表征。优化了聚苯乙烯微孔上仲胺共价连接的条件。在夹心杂交试验中，功能化载体用于共价结合DNA捕获探针以检测人巨细胞病毒。该试验的灵敏度与通过电磁辐射获得的市售表面Covalink-NH微孔板相比非常好。
