Glycobiology Research and Training Center, Department of Medicine, University of California, San Diego, La Jolla, California, United States.
Anal Chem. 2013 Apr 16;85(8):3864-70. doi: 10.1021/ac400320n. Epub 2013 Apr 2.
Current methods for analyzing sialic acid diversity in modifications and linkages require multistep processing, derivatization, and chromatographic analyses. We here report a single-step optical method for identification and quantification of different compositions of sialoglycans on glycoproteins and in serum. This was achieved by measuring and quantifying nanometal surface energy transfer (NSET) signals between quantum dots and gold nanoparticles bound to specific sialic acid binding proteins (SBPs) and sialic acid moieties, respectively. The biosensing process is based on the NSET turn-on by external sialic acid species that compete for binding to the SBPs. Selectivity of the biosensor toward sialoglycans can be designed to detect the total amount, glycosylation linkages (α2-6 vs α2-3), and modifications (9-O-acetyl and N-glycolyl groups) in the samples. This nanobiosensor is a prototype expected to achieve limits of the detection down to the micromolar range for high-throughput quantification and analysis of different compositions of sialoglycans present in biological or biomedical samples.
目前分析修饰和连接物中唾液酸多样性的方法需要多步处理、衍生化和色谱分析。我们在这里报告了一种用于鉴定和定量糖蛋白和血清中不同唾液酸聚糖组成的单步光学方法。这是通过测量和定量量子点和金纳米粒子之间的纳米金属表面能量转移(NSET)信号来实现的,分别与特定的唾液酸结合蛋白(SBPs)和唾液酸部分结合。生物传感过程基于外部唾液酸物种通过与 SBPs 竞争结合而引发的 NSET 开启。生物传感器对唾液酸聚糖的选择性可以设计为检测样品中的总量、糖基化连接(α2-6 与 α2-3)和修饰(9-O-乙酰基和 N-糖基化基团)。这种纳米生物传感器是一种原型,有望实现低至微摩尔范围的检测极限,用于高通量定量和分析生物或生物医学样品中存在的不同唾液酸聚糖组成。