title: Evaluation of the sulfo-succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate coupling chemistry for attachment of oligonucleotides to magnetic nanobeads.
abstract: The sulfo-SMCC (Succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate) coupling chemistry was evaluated for immobilization of oligonucleotides onto 130 nm sized magnetic nanobeads aimed for bio-detection in a magnetic readout assay. The chemistry was found to produce a high surface coverage of approximately 93 +/- 10 oligonucleotides per bead whereas stability tests showed that about 50% of the oligonucleotides detached from the bead surfaces after eight weeks of storage in a buffer solution. It was shown that bead aggregation prior to magnetic readout could be suppressed by incubating the samples at 70 degrees C for 30 min. The same temperature was also shown to be the most favorable for hybridization between the oligonucleotide functionalized beads and rolling circle amplified DNA molecules. This should simplify the heating procedure in a biosensor in which hybridization and magnetic readout is performed in the same compartment.
authors.affiliation: Department of Engineering Sciences, Uppsala University, The Angstrom Laboratory, Box 534, SE-751 21 Uppsala, Sweden.
authors.forename: Teresa Zardán Gómez
authors.initials: TZ
authors.lastname: de la Torre
authors.forename: David
authors.initials: D
authors.lastname: Herthnek
authors.forename: Harisha
authors.initials: H
authors.lastname: Ramachandraiah
authors.forename: Peter
authors.initials: P
authors.lastname: Svedlindh
authors.forename: Mats
authors.initials: M
authors.lastname: Nilsson
authors.forename: Maria
authors.initials: M
authors.lastname: Strømme
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D004247:DNA / Q000737:chemistry; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D008301:Maleimides / Q000737:chemistry*; D009693:Nucleic Acid Hybridization / Q000379:methods; D009841:Oligonucleotides / Q000737:chemistry*; D013499:Surface Properties; D013696:Temperature
chemical_list: D058185:Magnetite Nanoparticles; D008301:Maleimides; D009841:Oligonucleotides; C071675:sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate; D004247:DNA