title: ABCs of DNA aptamer and related assay development.
abstract: This review is intended to guide the novice in aptamer research and development to understand virtually all of the aptamer development options and currently available assay modalities. Aptamer development topics range from discussions of basic and advanced versions of Systematic Evolution of Ligands by EXponential Enrichment (SELEX) and SELEX variations involving incorporation of exotic unnatural nucleotides to expand library diversity for even greater aptamer affinity and specificity to improved next generation methods of DNA sequencing, screening and tracking aptamer development throughout the SELEX process and characterization of lead aptamer candidates. Aptamer assay development topics include descriptions of various colorimetric and fluorescent assays in microplates or on membranes including homogeneous beacon and multiplexed Fluorescence Resonance Energy Transfer (FRET) assays. Finally, a discussion of the potential for marketing successful aptamer-based assays or test kits is included.
authors.affiliation: Center for Biodesign and Diagnostics, Translational Health Science and Technology Institute, Faridabad, Haryana 121001, India; AptaBharat Innovation Private Limited, Translational Health Science and Technology Institute Incubator, Haryana 121001, India. Electronic address: tarun@thsti.res.in.
authors.forename: Tarun Kumar
authors.initials: TK
authors.lastname: Sharma
authors.affiliation: Operational Technologies Corporation, 4100 NW Loop 410, Suite, 230, San Antonio, TX 78229, USA.. Electronic address: john.bruno@otcorp.com.
authors.forename: John G
authors.initials: JG
authors.lastname: Bruno
authors.affiliation: Department of Biotechnology, All India Institute of Medical Sciences, New Delhi 110029, India.; Faculty of Pharmacy, Uttarakhand Technical University, Dehradun 248007, Uttarakhand, India.
authors.forename: Abhijeet
authors.initials: A
authors.lastname: Dhiman
mesh_terms: D052157:Aptamers, Nucleotide; D003124:Colorimetry; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D052156:SELEX Aptamer Technique
chemical_list: D052157:Aptamers, Nucleotide; D004247:DNA
keywords: Aptamer Linked Immunosorbent Assay (ALISA); Aptamer library; Aptamers; Colorimetric assay; FRET; Post-SELEX optimization; SELEX; Translation of aptamers; ssDNA generation