Amin Mohsin, Abdullah Badr M, Wylie Stephen R, Rowley-Neale Samuel J, Banks Craig E, Whitehead Kathryn A
Faculty of Engineering and Technology, Liverpool John Moores University, Liverpool L3 3AF, UK.
Faculty of Science and Engineering, Manchester Metropolitan University, Manchester M1 5GD, UK.
Nanomaterials (Basel). 2022 Dec 22;13(1):36. doi: 10.3390/nano13010036.
Cadaverine is a biomolecule of major healthcare importance in periodontal disease; however, current detection methods remain inefficient. The development of an enzyme biosensor for the detection of cadaverine may provide a cheap, rapid, point-of-care alternative to traditional measurement techniques. This work developed a screen-printed biosensor (SPE) with a diamine oxidase (DAO) and multi-walled carbon nanotube (MWCNT) functionalised electrode which enabled the detection of cadaverine via cyclic voltammetry and differential pulse voltammetry. The MWCNTs were functionalised with DAO using carbodiimide crosslinking with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) and -Hydroxysuccinimide (NHS), followed by direct covalent conjugation of the enzyme to amide bonds. Cyclic voltammetry results demonstrated a pair of distinct redox peaks for cadaverine with the C-MWCNT/DAO/EDC-NHS/GA SPE and no redox peaks using unmodified SPEs. Differential pulse voltammetry (DPV) was used to isolate the cadaverine oxidation peak and a linear concentration dependence was identified in the range of 3-150 µg/mL. The limit of detection of cadaverine using the C-MWCNT/DAO/EDC-NHS/GA SPE was 0.8 μg/mL, and the biosensor was also found to be effective when tested in artificial saliva which was used as a proof-of-concept model to increase the Technology Readiness Level (TRL) of this device. Thus, the development of a MWCNT based enzymatic biosensor for the voltammetric detection of cadaverine which was also active in the presence of artificial saliva was presented in this study.
尸胺是牙周疾病中具有重要医疗意义的生物分子;然而,目前的检测方法仍然效率低下。开发一种用于检测尸胺的酶生物传感器可能为传统测量技术提供一种廉价、快速的即时检测替代方法。这项工作开发了一种带有二胺氧化酶(DAO)和多壁碳纳米管(MWCNT)功能化电极的丝网印刷生物传感器(SPE),该传感器能够通过循环伏安法和差分脉冲伏安法检测尸胺。使用碳二亚胺与盐酸1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDC)和N-羟基琥珀酰亚胺(NHS)对MWCNT进行功能化,然后将酶直接共价偶联到酰胺键上。循环伏安法结果表明,C-MWCNT/DAO/EDC-NHS/GA SPE对尸胺有一对明显的氧化还原峰,而未修饰的SPE没有氧化还原峰。使用差分脉冲伏安法(DPV)分离尸胺氧化峰,并确定在3-150μg/mL范围内存在线性浓度依赖性。使用C-MWCNT/DAO/EDC-NHS/GA SPE检测尸胺的检测限为0.8μg/mL,并且当在用作概念验证模型的人工唾液中进行测试时,该生物传感器也被发现是有效的,以提高该设备的技术就绪水平(TRL)。因此,本研究提出了一种基于MWCNT的用于伏安法检测尸胺的酶生物传感器,该传感器在人工唾液存在下也具有活性。