Elsevier

Biotechnology Advances

Volume 37, Issue 1, January–February 2019, Pages 28-50
Biotechnology Advances

Research review paper
Three decades of nucleic acid aptamer technologies: Lessons learned, progress and opportunities on aptamer development

https://doi.org/10.1016/j.biotechadv.2018.11.001Get rights and content
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open access

Abstract

Aptamers are short single-stranded nucleic acid sequences capable of binding to target molecules in a way similar to antibodies. Due to various advantages such as prolonged shelf life, low batch to batch variation, low/no immunogenicity, freedom to incorporate chemical modification for enhanced stability and targeting capacity, aptamers quickly found their potential in diverse applications ranging from therapy, drug delivery, diagnosis, and functional genomics to bio-sensing. Aptamers are generated by a process called SELEX. However, the current overall success rate of SELEX is far from being satisfactory, and still presents a major obstacle for aptamer-based research and application. The need for an efficient selection strategy consisting of defined procedures to deal with a wide variety of targets is significantly important. In this work, by analyzing key aspects of SELEX including initial library design, target preparation, PCR optimization, and single strand DNA separation, we provide a comprehensive analysis of individual steps to facilitate researchers intending to develop personalized protocols to address many of the obstacles in SELEX. In addition, this review provides suggestions and opinions for future aptamer development procedures to address the concerns on key SELEX steps, and post-SELEX modifications.

Keywords

SELEX
Aptamer
SELEX library
Synthetic genetic polymers
Single-strand DNA preparation
PCR bias

Abbreviations

SELEX
Systematic Evolution of Ligands by Exponential enrichment
AEGIS
artificially expanded genetic information system
AFM
atomic force microscope
AON
antisense oligonucleotide
CMACS
continuous-flow magnetic activated chips
Ds
7-(2-thienyl) imidazo [4, 5-b] pyridine
EMSA
electrophoretic mobility shift assay
ECEEM
equilibrium capillary electrophoresis of equilibrium mixtures
ePCR
Emulsion PCR
FACS
fluorescence-activated cell sorter
FITC
Fluorescein isothiocyanate
Flu-Mag
Fluorescence-Magnetic
LNA
locked nucleic acid
MARAS
Magnetic-Assisted Rapid Aptamer Selection
NA
Nucleic acid
NRR
non-homologous random recombination
NGS
next generation sequencing
P
2-amino-8-(10-β-D-2-deoxyribofuranosyl)-imidazo[1,2-a]-1,3,5-triazin-4(8H)-one
PEG
polyethylene glycol
Px
2-nitro-4- propynylpyrrole
RAPID
RNA aptamer Isolation via dual-cycles
SweepCE
Sweeping capillary electrophoresis
SOMAmer
Slow off-rate modified aptamer
ssDNA
single-stranded DNA
TECS
target expressed on the cell surface
Thioaptamers
Thiophosphate-modified aptamers
UV-LEDIF
ultraviolet light-emitting diode-induced native fluorescence
Z
6-amino-5-nitro-3-(1'- β-D-2'-deoxyribofur-anosyl)-2(1H)-pyridone
μFFE
Micro Free Flow Electrophoresis
2′-FANA
2'-fluoroarabino nucleic acid

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1

These authors contributed equally to this work.