Point-of-Care Analyte Quantification and Digital Readout via Lysate-Based Cell-Free Biosensors Interfaced with Personal Glucose Monitors.
Point-of-Care Analyte Quantification and Digital Readout via Lysate-Based Cell-Free Biosensors Interfaced with Personal Glucose Monitors.
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DOI：
10.1021/acssynbio.1c00282
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发表时间：
2021-11-19
影响因子：
4.7
通讯作者：
Styczynski, Mark P.
中科院分区：
文献类型：
--
作者：
Zhang, Yan;Steppe, Paige L.;Kazman, Maxwell W.;Styczynski, Mark P.
基于无细胞系统的现场可部署诊断技术有了很大进步，但目标分析物的现场量化仍然是一个挑战。在这里，我们展示了基于大肠杆菌裂解物的无细胞生物传感器与个人血糖监测仪(PGM)相结合可以实现现场分析物定量，并具有直接重新配置不同类型分析物的潜力。我们发现分析物响应的转录和翻译调节因子可以调节报告酶β-半乳糖苷酶的产生，该酶反过来将乳糖转化为葡萄糖用于PGM的定量。因为糖酵解在裂解物中很活跃，很容易消耗转化的葡萄糖，所以我们分离了酶的产生和葡萄糖的转化，以增加终点信号输出。然而，这种裂解产物代谢确实允许一锅法去除复杂样品(如人血清)中存在的葡萄糖，而不会混淆目标定量。综上所述，我们表明，将基于裂解物的无细胞生物传感器与PGMS相结合，可以在需要时进行可访问的目标检测和量化。
Field-deployable diagnostics based on cell-free systems have advanced greatly, but on-site quantification of target analytes remains a challenge. Here we demonstrate that Escherichia coli lysate-based cell-free biosensors coupled to a personal glucose monitor (PGM) can enable on-site analyte quantification, with the potential for straightforward reconfigurability to diverse types of analytes. We show that analyte-responsive regulators of transcription and translation can modulate production of the reporter enzyme β-galactosidase, which in turn converts lactose into glucose for PGM quantification. Because glycolysis is active in the lysate and would readily deplete converted glucose, we decoupled enzyme production and glucose conversion to increase endpoint signal output. This lysate metabolism did, however, allow for one-pot removal of glucose present in complex samples (like human serum) without confounding target quantification. Taken together, we show that integrating lysate-based cell-free biosensors with PGMs enables accessible target detection and quantification at the point of need.
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影响因子：
64.5
作者：
Pardee K;Green AA;Ferrante T;Cameron DE;DaleyKeyser A;Yin P;Collins JJ
通讯作者：
Collins JJ
DOI：
10.1111/nyas.12069
发表时间：
2013-01-01
期刊：
ANNALS MEETING REPORTS
影响因子：
--
作者：
Combs, Gerald F., Jr.;Trumbo, Paula R.;Raiten, Daniel J.
通讯作者：
Raiten, Daniel J.
影响因子：
4.6
作者：
Kwon YC;Jewett MC
通讯作者：
Jewett MC
影响因子：
8.4
作者：
Caschera, Filippo;Noireaux, Vincent
通讯作者：
Noireaux, Vincent
影响因子：
3.2
作者：
Karim, Ashty S.;Rasor, Blake J.;Jewett, Michael C.
通讯作者：
Jewett, Michael C.