适体
指数富集配体系统进化
表面等离子共振
SELEX适体技术
微流控
纳米技术
化学
核糖核酸
材料科学
生物物理学
生物
分子生物学
纳米颗粒
基因
生物化学
作者
Eric Dausse,Aurélien Barré,Ahissan Aimé,Alexis Groppi,Alain Rico,Chrysanthi Ainali,Gilmar F. Salgado,William Palau,Emilie Daguerre,Macha Nikolski,Jean‐Jacques Toulmé,Carmelo Di Primo
标识
DOI:10.1016/j.bios.2016.02.003
摘要
A surface plasmon resonance (SPR)-based SELEX approach has been used to raise RNA aptamers against a structured RNA, derived from XBP1 pre-mRNA, that folds as two contiguous hairpins. Thanks to the design of the internal microfluidic cartridge of the instrument, the selection was performed during the dissociation phase of the SPR analysis by recovering the aptamer candidates directly from the target immobilized onto the sensor chip surface. The evaluation of the pools was performed by SPR, simultaneously, during the association phase, each time the amplified and transcribed candidates were injected over the immobilized target. SPR coupled with SELEX from the first to the last round allowed identifying RNA aptamers that formed highly stable loop-loop complexes (KD equal to 8nM) with the hairpin located on the 5' side of the target. High throughput sequencing of two key rounds confirmed the evolution observed by SPR and also revealed the selection of hairpins displaying a loop not fully complementary to the loop of its target. These candidates were selected mainly because they bound 79 times faster to the target than those having a complementary loop. SELEX coupled with SPR is expected to speed up the selection process because selection and evaluation are performed simultaneously.
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