title: Selection of Aptamers for Metabolite Sensing and Construction of Optical Nanosensors.
abstract: Optical nanosensors are based on particles with diameters from 20 to 200 nm containing sensory elements. The latter are comprised of one or more signaling molecules and one or more references, which allow measurements to be ratiometric and hence independent on the amount of sensor. The signaling molecules may range from simple ion-binding fluorophores, e.g., pH-sensitive dyes, to complex biochemical assays. Aptamers are ideal for use in nanosensors because they are relatively easy to modify chemically and hence to transform into signaling molecules, and their binding affinities may be fine-tuned to a desired measuring range in the selection process. Here we first describe the selection of metabolite binding aptamers, how they are transformed into signaling molecules using a molecular beacon construct and then how they are inserted into nanoparticles. Finally, we briefly describe how the sensors are calibrated before inserted into cells to measure metabolite concentration in real time. As examples we present aptamers binding to key metabolites in cells: ATP and fructose 1, 6-bisphosphate (FBP).
authors.affiliation: Institute of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230, Odense, Denmark.|LIMES Institute, University of Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany.
authors.forename: Yi
authors.initials: Y
authors.lastname: Long
authors.affiliation: LIMES Institute, University of Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Pfeiffer
authors.affiliation: LIMES Institute, University of Bonn, Gerhard-Domagk-Str. 1, 53121, Bonn, Germany.
authors.forename: Günter
authors.initials: G
authors.lastname: Mayer
authors.affiliation: Institute of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230, Odense, Denmark.|Lundbeckfonden Center of Excellence NanoCAN, University of Southern Denmark, Campusvej 55, 5230, Odense, Denmark.
authors.forename: Tine Daa
authors.initials: TD
authors.lastname: Schrøder
authors.affiliation: Department of Medical Biology, School of Medicine, Istanbul Kemerburgaz University, 34217, Istanbul, Turkey.
authors.forename: Veli Cengiz
authors.initials: VC
authors.lastname: Özalp
authors.affiliation: Institute of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230, Odense, Denmark. lfo@bmb.sdu.dk.
authors.forename: Lars Folke
authors.initials: LF
authors.lastname: Olsen
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D052157:Aptamers, Nucleotide / Q000737:chemistry; D015374:Biosensing Techniques; D002138:Calibration; D005635:Fructosediphosphates / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods*; D052156:SELEX Aptamer Technique
chemical_list: D052157:Aptamers, Nucleotide; D005635:Fructosediphosphates; D000255:Adenosine Triphosphate; C029063:fructose-1,6-diphosphate
keywords: 6-Bisphosphate; ATP; Aptamers; Calibration; Fructose 1; Nanosensor; Selection