Creating advanced multifunctional biosensors with surface enzymatic transformations
Creating advanced multifunctional biosensors with surface enzymatic transformations
复制标题
DOI：
10.1021/la060223o
复制
发表时间：
2006-06-06
期刊：
影响因子：
3.9
通讯作者：
Corn, Robert M.
中科院分区：
文献类型：
--
作者：
Lee, Hye Jin;Wark, Alastair W.;Corn, Robert M.
本文总结了我们最近在生物聚合物微阵列表面酶化学和生物亲和力相互作用的耦合方面所做的工作，以创建具有更高选择性和灵敏度的多路生物传感器。利用表面等离子体共振成像(SPRI)和表面等离子体荧光光谱(SPFS)这两种表面敏感技术，实时检测表面酶的转化。展示了三个新的表面生物亲和力/表面酶耦合过程的具体例子：(I)结合RNA微阵列使用酶核糖核酸酶H(RNaseH)的表面酶扩增方法，该方法允许在1fM的浓度下超灵敏地直接检测基因组DNA，而不需要标记或PCR扩增；(Ii)使用RNA-DNA连接化学来从单链DNA微阵列创建可再生的RNA微阵列；以及(Iii)应用T7RNA聚合酶从双链DNA微阵列元件上复制RNA。此外，一个简单而强大的理论框架，包括酶吸附和表面酶动力学的贡献，被用来定量表面酶的反应性。该模型被成功地应用于寡核苷酸芯片上核糖核酸酶H和外切核酸酶III(Exo III)表面水解反应的SPRI和SPFS测量。
This paper summarizes our recent work on the coupling of surface enzyme chemistry and bioaffinity interactions on biopolymer microarrays for the creation of multiplexed biosensors with enhanced selectivity and sensitivity. The surface sensitive techniques of surface plasmon resonance imaging (SPRI) and surface plasmon fluorescence spectroscopy (SPFS) are used to detect the surface enzymatic transformations in real time. Three specific examples of novel coupled surface bioaffinity/surface enzymatic processes are demonstrated: (i) a surface enzymatic amplification method utilizing the enzyme ribonuclease H (RNase H) in conjunction with RNA microarrays that permits the ultrasensitive direct detection of genomic DNA at a concentration of 1 fM without labeling or PCR amplification, (ii) the use of RNA-DNA ligation chemistry to create renewable RNA microarrays from single stranded DNA microarrays, and (iii) the application of T7 RNA polymerase for the on-chip replication of RNA from double stranded DNA microarray elements. In addition, a simple yet powerful theoretical framework that includes the contributions of both enzyme adsorption and surface enzyme kinetics is used to quantitate surface enzyme reactivity. This model is successfully applied to SPRI and SPFS measurements of surface hydrolysis reactions of RNase H and Exonuclease III (Exo III) on oligonucleotide microarrays.