title: Integrated Microfluidic Isolation of Aptamers Using Electrophoretic Oligonucleotide Manipulation.
abstract: We present a microfluidic approach to integrated isolation of DNA aptamers via systematic evolution of ligands by exponential enrichment (SELEX). The approach employs a microbead-based protocol for the processes of affinity selection and amplification of target-binding oligonucleotides, and an electrophoretic DNA manipulation scheme for the coupling of these processes, which are required to occur in different buffers. This achieves the full microfluidic integration of SELEX, thereby enabling highly efficient isolation of aptamers in drastically reduced times and with minimized consumption of biological material. The approach as such also offers broad target applicability by allowing selection of aptamers with respect to targets that are either surface-immobilized or solution-borne, potentially allowing aptamers to be developed as readily available affinity reagents for a wide range of targets. We demonstrate the utility of this approach on two different procedures, respectively for isolating aptamers against a surface-immobilized protein (immunoglobulin E) and a solution-phase small molecule (bisboronic acid in the presence of glucose). In both cases aptamer candidates were isolated in three rounds of SELEX within a total process time of approximately 10 hours.
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, New York 10027, United States.
authors.forename: Jinho
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, New York 10027, United States.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Olsen
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, New York 10027, United States.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, New York 10027, United States.
authors.forename: John P
authors.initials: JP
authors.lastname: Hilton
authors.affiliation: Division of Clinical Pharmacology and Experimental Therapeutics, Department of Medicine, Columbia University, Columbia University, New York, NY 10032, United States.
authors.forename: Kyung-Ae
authors.initials: KA
authors.lastname: Yang
authors.affiliation: Division of Nanobiomedicine, Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, Jiangsu 215123, China.
authors.forename: Renjun
authors.initials: R
authors.lastname: Pei
authors.affiliation: Division of Clinical Pharmacology and Experimental Therapeutics, Department of Medicine, Columbia University, Columbia University, New York, NY 10032, United States.
authors.forename: Milan N
authors.initials: MN
authors.lastname: Stojanovic
authors.affiliation: Department of Mechanical Engineering, Columbia University, New York, New York 10027, United States.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Lin
mesh_terms: D052157:Aptamers, Nucleotide / Q000302:isolation & purification*; D001897:Boronic Acids / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods*; D007073:Immunoglobulin E / Q000378:metabolism; D044085:Microfluidics / Q000379:methods*; D052156:SELEX Aptamer Technique
chemical_list: D052157:Aptamers, Nucleotide; D001897:Boronic Acids; D007073:Immunoglobulin E