Creating advanced multifunctional biosensors with surface enzymatic transformations
Lee, H.J. and Wark, A.W. and Corn, R.M. (2006) Creating advanced multifunctional biosensors with surface enzymatic transformations. Langmuir, 22 (12). pp. 5241-5250. ISSN 0743-7463 (http://pubs.acs.org/doi/abs/10.1021/la060223o)
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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 RNADNA ligation chemistry to create renewable RNA microarrays from single stranded DNA microarrays, and (iii) the application of T7RNApolymerase for the on-chip replication ofRNAfrom double strandedDNAmicroarray 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
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Item type: Article ID code: 9935 Dates: DateEvent6 June 2006PublishedSubjects: Science > Chemistry Department: Faculty of Science > Pure and Applied Chemistry Depositing user: Strathprints Administrator Date deposited: 25 Nov 2011 14:41 Last modified: 24 Apr 2024 04:59 URI: https://strathprints.strath.ac.uk/id/eprint/9935