【2h】

Micromagnetic selection of aptamers in microfluidic channels

机译:微流体通道中适体的微磁选择

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摘要

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.
机译:适体是已经在体外选择以高亲和力和特异性结合其分子靶标的核酸分子。通常,通过指数富集(SELEX)过程对配体的系统进化被用于分离特定的高亲和力适体。但是,SELEX是一个迭代过程,需要多轮选择和放大,这需要大量的时间和精力。在这里,我们将基于磁珠的SELEX工艺与微流体技术相结合,描述了一种适配子发现系统,该系统快速,高效,可自动化且适用于广泛的目标。我们的微流体SELEX(M-SELEX)方法利用了在微尺度上发生的许多独特现象,并实施了一种设计,使其能够精确地操纵少量的珠子并快速分离出高亲和力的适体。作为展示M-SELEX过程效率的模型,我们在这里描述了单轮选择后DNA适体的分离,该适体紧密结合重组A型肉毒杆菌神经毒素轻链(具有低纳摩尔解离常数)。

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