A Personal Glucose Meter for Label-Free and Washing-Free Biomolecular Detection
A Personal Glucose Meter for Label-Free and Washing-Free Biomolecular Detection
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DOI：
10.1021/acs.analchem.8b02014
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发表时间：
2018-10-02
影响因子：
7.4
通讯作者：
Park, Hyun Gyu
中科院分区：
文献类型：
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作者：
Ahn, Jun Ki;Kim, Hyo Yong;Park, Hyun Gyu
我们开发了一种使用个人血糖仪(PGM)的免标记、免洗涤的生物分子检测方法。以三磷酸腺苷为模型靶点，采用己糖激酶和丙酮酸激酶促进的级联酶反应，将三磷酸腺苷的量连接到手持式PGM可检测到的葡萄糖上。原则上，靶标三磷酸腺苷的存在使己糖激酶能够通过向葡萄糖提供磷酸基团来催化葡萄糖转化为葡萄糖6-磷酸，从而葡萄糖的量与三磷酸腺苷的量成比例地减少。此外，己糖酶催化反应后产生的腺苷5‘-二磷酸(ADP)被丙酮酸激酶酶还原为ATP。再生的ATP再次被添加以催化多轮级联酶反应，从而导致信号放大。因此，葡萄糖含量的变化与三磷酸腺苷含量成反比，简单地通过手持PGM来测量。通过使用该策略，我们成功地将ATP低至49 nM，即使在真实样品中，如自来水、人血清和牛尿，也具有高选择性。重要的是，所开发的系统不需要昂贵的改装和清洗步骤，而是使用商业化的PGM操作方便，这将为开发简单且具有成本效益的传感平台铺平道路。
We developed a label-free and washing-free method for biomolecular detection using a personal glucose meter (PGM). ATP was selected as a model target, and cascade enzymatic reactions promoted by hexokinase and pyruvate kinase were adopted to link the amount of ATP to glucose that is detectable by a hand-held PGM. In principle, the presence of target ATP enables hexokinase to catalyze the conversion of glucose to glucose 6-phosphate by providing a phosphate group to glucose, and thus the amount of glucose is decreased in proportion to the amount of ATP. In addition, adenosine 5'-diphosphate (ADP), which is generated after hexokinase-catalyzed enzymatic reaction, is recovered to ATP by a pyruvate kinase enzyme. The regenerated ATP is again supplemented to catalyze multiple rounds of cascade enzymatic reactions, leading to signal amplification. As a result, the change of glucose amount that is inversely proportional to ATP amount is simply measured by a hand-held PGM. By employing this strategy, we successfully determined ATP down to 49 nM with high selectivity even in real samples such as tap water, human serum, and bovine urine. Importantly, the developed system does not require expensive modification and washing steps but is conveniently operated with a commercially available PGM, which would pave the way for the development of a simple and cost-effective sensing platform.