Polyak Boris, Geresh Shimona, Marks Robert S
The Institute for Applied Biosciences and Department of Biotechnology Engineering, Ben-Gurion University of the Negev, PO Box 653, Beer Sheva 84105, Israel.
Biomacromolecules. 2004 Mar-Apr;5(2):389-96. doi: 10.1021/bm034454a.
Biotin was covalently coupled with alginate in an aqueous-phase reaction by means of carbodiimide-mediated activation chemistry to provide a biotin-alginate conjugate for subsequent use in biosensor applications. The synthetic procedure was optimized with respect to pH of the reaction medium (pH 6.0), the degree of uronic acid activation (20%), and the order of addition of the reagents. The biotin-alginate conjugate was characterized by titration with 2-anilinonaphthalene-6-sulfonic acid (2,6-ANS), 4-hydroxyazobene-2'-carboxylic acid (HABA) and by an HPSEC-MALLS analytical method as well as by FTIR and 13C NMR spectroscopy. As a compromise between the need for a high percent of molar modification of the alginate, on one hand, and sufficient gelling capability, on the other hand, an optimal modification of 10-13% of biotin-alginate was used. The new biotin-alginate conjugate was used for the encapsulation of bioluminescent reporter cells into microspheres. A biosensor was prepared by conjugating these biotinylated alginate microspheres to the surface of a streptavidin-coated optical fiber, and the performance of the biosensor was demonstrated in the determination of the antibiotic, mitomycin C as a model toxin.
通过碳二亚胺介导的活化化学方法，在水相反应中将生物素与海藻酸盐共价偶联，以提供生物素 - 海藻酸盐共轭物，用于后续的生物传感器应用。合成过程针对反应介质的pH值（pH 6.0）、糖醛酸活化程度（20%）和试剂添加顺序进行了优化。生物素 - 海藻酸盐共轭物通过用2 - 苯胺基萘 - 6 - 磺酸（2,6 - ANS）、4 - 羟基偶氮苯 - 2'- 羧酸（HABA）滴定，以及HPSEC - MALLS分析方法、FTIR和13C NMR光谱进行表征。一方面，在海藻酸盐高摩尔修饰百分比的需求与另一方面足够的凝胶化能力之间取得平衡，使用了10 - 13%的生物素 - 海藻酸盐最佳修饰。新的生物素 - 海藻酸盐共轭物用于将生物发光报告细胞封装到微球中。通过将这些生物素化的海藻酸盐微球与链霉亲和素包被的光纤表面偶联制备了生物传感器，并在测定抗生素丝裂霉素C作为模型毒素时展示了该生物传感器的性能。
