title: Aptamer-based trapping of phytosphingosine in urine samples.
abstract: Usually, small molecules like single metabolites used in clinical diagnostic can be quantified by instrumental approaches like LC-MS or bioanalytical techniques using antibodies or aptamers as selective receptors. The present work comprises the generation of aptamers with an affinity towards the medically relevant metabolite phytosphingosine via the previously reported just in time-Selection approach (Hünniger et al., 2014). The whole approach could be seen as a proof of concept to extend the existing just in time-Selection protocol for selection towards small molecules with dissociation constants in the low nanomolar range. Moreover it is conceivable that the shown methods could be quickly adapted to further scopes. Aptamers could be applied for clean-up or concentration processes prior to further analysis. As an example, we used the selected aptamers towards phytosphingosine bound to magnetic particles for affinity enrichment in both selection buffer and urine samples. As an outcome, enrichment factors of up to 9-fold (selection buffer)/4-fold (urine samples) were achieved by this approach.
authors.affiliation: HAMBURG SCHOOL OF FOOD SCIENCE; Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.
authors.forename: Christin
authors.initials: C
authors.lastname: Fischer
authors.affiliation: HAMBURG SCHOOL OF FOOD SCIENCE; Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Klockmann
authors.affiliation: HAMBURG SCHOOL OF FOOD SCIENCE; Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.
authors.forename: Hauke
authors.initials: H
authors.lastname: Wessels
authors.affiliation: HAMBURG SCHOOL OF FOOD SCIENCE; Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Hünniger
authors.affiliation: HAMBURG SCHOOL OF FOOD SCIENCE; Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.
authors.forename: Jil
authors.initials: J
authors.lastname: Schrader
authors.affiliation: HAMBURG SCHOOL OF FOOD SCIENCE; Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany.
authors.forename: Angelika
authors.initials: A
authors.lastname: Paschke-Kratzin
authors.affiliation: HAMBURG SCHOOL OF FOOD SCIENCE; Institute of Food Chemistry, University of Hamburg, Grindelallee 117, 20146 Hamburg, Germany. Electronic address: Markus.Fischer@uni-hamburg.de.
authors.forename: Markus
authors.initials: M
authors.lastname: Fischer
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015415:Biomarkers / Q000652:urine; D002853:Chromatography, Liquid; D006801:Humans; D057230:Limit of Detection; D059346:Magnets; D021241:Spectrometry, Mass, Electrospray Ionization; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000652:urine; D053719:Tandem Mass Spectrometry
chemical_list: D052157:Aptamers, Nucleotide; D015415:Biomarkers; C012491:phytosphingosine; D013110:Sphingosine
keywords: Aptamer; Metabolite; Small molecule; Trapping; Urine; just in time-Selection