Optimization of Vancomycin Aptamer Sequence Length Increases the Sensitivity of Electrochemical, Aptamer-Based Sensors In Vivo.
Optimization of Vancomycin Aptamer Sequence Length Increases the Sensitivity of Electrochemical, Aptamer-Based Sensors In Vivo.
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实用万古霉素适体序列长度的优化增加了体内电化学，基于适体的传感器的灵敏度。
DOI：
10.1021/acssensors.2c01910
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发表时间：
2022-12-23
期刊：
影响因子：
8.9
通讯作者：
Arroyo-Curras, Netzahualcoyotl
中科院分区：
文献类型：
--
作者：
Shaver, Alexander;Mahlum, J. D.;Scida, Karen;Johnston, Melanie L.;Pellitero, Miguel Aller;Wu, Yao;Carr, Gregory V.;Arroyo-Curras, Netzahualcoyotl
鉴于万古霉素的治疗窗较窄，在临床上测量血清万古霉素水平对于优化剂量至关重要。目前定量血清万古霉素水平的方法是基于免疫测定法，这是一种需要对患者样本进行大量处理的多步骤方法。作为替代方案，开发了万古霉素结合电化学、基于适体的传感器（E-AB），以简化万古霉素监测的工作流程。E-AB能够即时测量血清万古霉素浓度，无需样本稀释或其他处理步骤。然而，最初报告的万古霉素结合E-AB的解离常数为45 μ M，比在患者中测量的万古霉素推荐谷浓度高约1个数量级。这种有限的灵敏度阻碍了E-AB准确支持万古霉素监测的能力。为了克服这个问题，我们在这里试图优化万古霉素结合适体序列的长度，以在E-AB平台中实现更宽的动态范围。我们的研究结果表明，通过等温量热法和E-AB校准在未稀释的血清中，上级的亲和力和接近相等的传感器增益在体外可以实现使用一个碱基对更长的适配体比原来报道的截短序列。我们通过监测静脉给药后活小鼠脑皮质中万古霉素水平来验证体内结合亲和力改善的影响。虽然原始序列无法从基线噪声中分辨万古霉素浓度（SNR = 1.03），但我们新报告的序列在相同剂量下提供了1.62的SNR。
The measurement of serum vancomycin levels at the clinic is critical to optimizing dosing given the narrow therapeutic window of this antibiotic. Current approaches to quantitate serum vancomycin levels are based on immunoassays, which are multistep methods requiring extensive processing of patient samples. As an alternative, vancomycin-binding electrochemical, aptamer-based sensors (E-ABs) were developed to simplify the workflow of vancomycin monitoring. E-ABs enable the instantaneous measurement of serum vancomycin concentrations without the need for sample dilution or other processing steps. However, the originally reported vancomycin-binding E-ABs had a dissociation constant of 45 μM, which is approximately 1 order of magnitude higher than the recommended trough concentrations of vancomycin measured in patients. This limited sensitivity hinders the ability of E-ABs to accurately support vancomycin monitoring. To overcome this problem, here we sought to optimize the length of the vancomycin-binding aptamer sequence to enable a broader dynamic range in the E-AB platform. Our results demonstrate, via isothermal calorimetry and E-AB calibrations in undiluted serum, that superior affinity and near-equal sensor gain in vitro can be achieved using a one-base-pair-longer aptamer than the truncated sequence originally reported. We validate the impact of the improved binding affinity in vivo by monitoring vancomycin levels in the brain cortex of live mice following intravenous administration. While the original sequence fails to resolve vancomycin concentrations from baseline noise (SNR = 1.03), our newly reported sequence provides an SNR of 1.62 at the same dose.
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影响因子：
9.5
作者：
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通讯作者：
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影响因子：
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作者：
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影响因子：
11.8
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影响因子：
7.4
作者：
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通讯作者：
Plaxco, Kevin W.
影响因子：
9.5
作者：
Shaver, Alexander;Curtis, Samuel D.;Arroyo-Curras, Netzahualcoyotl
通讯作者：
Arroyo-Curras, Netzahualcoyotl