Analytical and Bioanalytical Chemistry ( IF 4.3 ) Pub Date : 2018-10-01 , DOI: 10.1007/s00216-018-1398-7 Yucheng Liu , Bao Wang , Xinghu Ji , Zhike He
Multi-protein (or enzyme) conjugates play a vital role in biosensing due to the integrated function of each component, such as biological recognition and signal amplification. In this work, a green self-assembled method for the synthesis of multi-functional protein-enzyme nanoflowers has been developed, in which no chemical modification and coupling reaction is needed to fabricate the fluorescent signal probe. The self-assembled protein-enzyme conjugates streptavidin (SA) -β-galactosidase (β-Gal)-CaHPO4 nanoflowers load sufficient enzymes without damaging their activity, which meets the requirements of signal tags for biosensing. Through integrated multi-function of biorecognition (SA) and signal amplification (β-Gal), the SA-β-Gal-CaHPO4 hybrid nanoflower-based fluorescent sensor exhibited an ultrasensitive detection of protein biomarker alpha-fetoprotein (AFP), with limits of detection at the fM level. The presented self-assembled strategy can be extensively applied to develop on-demand protein-enzyme conjugates according to the specific requirements in a variety of applications including biosensors, bioimaging, and biomedicine.
中文翻译:
自组装的基于蛋白质酶纳米花的荧光传感技术,用于蛋白质生物标志物
由于每种成分的整合功能(例如生物识别和信号放大),多蛋白(或酶)结合物在生物传感中起着至关重要的作用。在这项工作中,已开发出一种绿色的自组装方法,用于合成多功能蛋白质酶纳米花,该方法不需要化学修饰和偶联反应即可制备荧光信号探针。自组装的蛋白质-酶结合物链霉亲和素(SA)-β-半乳糖苷酶(β-Gal)-CaHPO 4纳米花可装载足够的酶而不会破坏其活性,这满足了生物传感信号标签的要求。通过生物识别(SA)的集成多功能和信号放大(β-GAL),所述SA-β-Gal一使用CaHPO 4基于杂交纳米花的荧光传感器表现出对蛋白质生物标志物甲胎蛋白(AFP)的超灵敏检测,检测限在fM级别。提出的自组装策略可根据各种应用的具体要求,广泛应用于开发按需蛋白质-酶结合物,包括生物传感器,生物成像和生物医学。