Nanomaterials functionalized signal-ON/OFF electrogenerated chemiluminescence biosensor for quantization of trypsin based on target-induced cleavage of peptide
Shuo Kang, Ronghan Qu, Yuhong Duan, Jiale Yang, Fen Ma
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引用次数: 0
Abstract
Trypsin (TPN) is an important proteolytic enzyme in the digestive system and its abnormal levels are indicative of some pancreatic diseases. As an endopeptidase, TPN can cleave substrate peptide mainly by catalyzing the hydrolysis of the carboxyl side peptide bond of lysine (K) or arginine (R) residues. Based on this hydrolysis cleavage effect, two kinds of nanomaterials functionalized electrogenerated chemiluminescence (ECL) biosensors for the determination of TPN were designed as follows: A signal-ON ECL biosensor was fabricated by attaching substrate peptide (HWR*GWVC, “*” representing the cleavage site, abbreviated as HGC) labeled with ferrocene carboxylic acid (as quencher) onto the surface of NH2-MIL-53(Al) film which was incorporated with ECL emitting species (bis (2,2′-bipyridine)-4′-methyl-4-carboxybipyridine-ruthenium (N-succinimidylester)-bis (hexafluorophosphate) (Ru complex)). The presence of tryptic cleavage event can eventually lead the Fc complex to leave the electrode and results in the increase of the ECL intensity. HGC labeled with Ru-Ti3C2Tx-AuNP complex was used as capture probe and signal probe, which was attached onto NH2-MIL-53(Al) film modified glassy carbon electrode. A signal-OFF ECL biosensor was built as described above. The presence of TPN lead the Ru-Ti3C2Tx-AuNPs to leave the electrode, which resulted in the decreasing ECL intensity. As expected, the fabricated ECL biosensing methods provide excellent sensitivity and selectivity toward the TPN activity. Thus, this strategy shows great potential application in the clinic for diagnosis of TPN-indicating diseases as well as the screening of TPN inhibitor-based anti-cancer drugs.
胰蛋白酶（TPN）是消化系统中一种重要的蛋白水解酶，其水平异常可提示某些胰腺疾病。作为一种内肽酶，TPN主要通过催化水解赖氨酸(K)或精氨酸(R)残基的羧基侧肽键来裂解底物肽。基于这种水解裂解效应，设计了两种纳米材料功能化的电化学发光（ECL）生物传感器来测定TPN：将二茂铁羧酸（作为猝灭剂）标记的底物肽（HWR*GWVC，“*”表示裂解位点，缩写为HGC）附着在NH2-MIL-53（Al）膜表面，该膜与ECL发射物质(bis（2,2′-联吡啶）-4′-甲基-4-羧基联吡啶-钌（n -琥珀酰亚胺酯）-bis（六氟磷酸）（Ru配合物）结合，制成信号- on ECL生物传感器。色氨酸裂解事件的存在最终会导致Fc复合物离开电极，导致ECL强度增加。采用Ru-Ti3C2Tx-AuNP配合物标记的HGC作为捕获探针和信号探针，连接到NH2-MIL-53（Al）膜修饰的玻碳电极上。如上所述，构建了信号关闭ECL生物传感器。TPN的存在导致Ru-Ti3C2Tx-AuNPs离开电极，导致ECL强度降低。正如预期的那样，制备的ECL生物传感方法对TPN活性具有良好的灵敏度和选择性。因此，该策略在临床诊断TPN指示疾病以及筛选基于TPN抑制剂的抗癌药物方面具有很大的应用潜力。图形抽象
期刊介绍：
As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.
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