Micromagnetic selection of aptamers in microfluidic channels.
Aptamers are nucleic acid molecules that have been selected in vitro to bind to their molecular targets with high affinity and specificity. Typically, the systematic evolution of ligands by exponential enrichment (SELEX) process is used for the isolation of specific, high-affinity aptamers. SELEX, however, is an iterative process requiring multiple rounds of selection and amplification that demand significant time and labor. Here, we describe an aptamer discovery system that is rapid, highly efficient, automatable, and applicable to a wide range of targets, based on the integration of magnetic bead-based SELEX process with microfluidics technology. Our microfluidic SELEX (M-SELEX) method exploits a number of unique phenomena that occur at the microscale and implements a design that enables it to manipulate small numbers of beads precisely and isolate high-affinity aptamers rapidly. As a model to demonstrate the efficiency of the M-SELEX process, we describe here the isolation of DNA aptamers that tightly bind to the light chain of recombinant Botulinum neurotoxin type A (with low-nanomolar dissociation constant) after a single round of selection.
Lou
Xinhui
X
Department of Materials, University of California, Santa Barbara, CA 93106, USA.
Qian
Jiangrong
J
Xiao
Yi
Y
Viel
Lisan
L
Gerdon
Aren E
AE
Lagally
Eric T
ET
Atzberger
Paul
P
Tarasow
Theodore M
TM
Heeger
Alan J
AJ
Soh
H Tom
HT
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000302:isolation & purification*; D030561:Databases, Nucleic Acid; D008280:Magnetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D009690:Nucleic Acid Conformation
D052157:Aptamers, Nucleotide