TY - JOUR
T1 - An integrated microfluidic SELEX approach using combined electrokinetic and hydrodynamic manipulation
AU - Olsen, Timothy
AU - Zhu, Jing
AU - Kim, Jinho
AU - Pei, Renjun
AU - Stojanovic, Milan N.
AU - Lin, Qiao
N1 - Publisher Copyright:
© 2016 Society for Laboratory.
PY - 2017/2
Y1 - 2017/2
N2 - This article presents a microfluidic approach for the integration of the process of aptamer selection via systematic evolution of ligands by exponential enrichment (SELEX). The approach employs bead-based biochemical reactions in which affinityselected target-binding oligonucleotides are electrokinetically transferred for amplification, while the amplification product is transferred back for affinity selection via pressure-driven fluid flow. The hybrid approach simplifies the device design and operation procedures by reduced pressure-driven flow control requirements and avoids the potentially deleterious exposure of targets to electric fields prior to and during affinity selection. In addition, bead-based reactions are used to achieve the on-chip coupling of affinity selection and amplification of target-binding oligonucleotides, thereby realizing on-chip loop closure and integration of the entire SELEX process without requiring offline procedures. The microfluidic approach is thus capable of closed-loop, multiround aptamer enrichment as demonstrated by selection of DNA aptamers against the protein immunoglobulin E with high affinity (KD = 12 nM) in a rapid manner (4 rounds in approximately 10 h).
AB - This article presents a microfluidic approach for the integration of the process of aptamer selection via systematic evolution of ligands by exponential enrichment (SELEX). The approach employs bead-based biochemical reactions in which affinityselected target-binding oligonucleotides are electrokinetically transferred for amplification, while the amplification product is transferred back for affinity selection via pressure-driven fluid flow. The hybrid approach simplifies the device design and operation procedures by reduced pressure-driven flow control requirements and avoids the potentially deleterious exposure of targets to electric fields prior to and during affinity selection. In addition, bead-based reactions are used to achieve the on-chip coupling of affinity selection and amplification of target-binding oligonucleotides, thereby realizing on-chip loop closure and integration of the entire SELEX process without requiring offline procedures. The microfluidic approach is thus capable of closed-loop, multiround aptamer enrichment as demonstrated by selection of DNA aptamers against the protein immunoglobulin E with high affinity (KD = 12 nM) in a rapid manner (4 rounds in approximately 10 h).
KW - Lab-on-a-chip
KW - Microfluidics
KW - Molecular biology
KW - Systems
UR - http://www.scopus.com/inward/record.url?scp=85014488172&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85014488172&partnerID=8YFLogxK
U2 - 10.1177/2211068216659255
DO - 10.1177/2211068216659255
M3 - Article
C2 - 27418370
AN - SCOPUS:85014488172
SN - 2472-6303
VL - 22
SP - 63
EP - 72
JO - SLAS Technology
JF - SLAS Technology
IS - 1
ER -