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基于金纳米粒子复合石墨化介孔碳和载阿霉素空心金纳米球的高灵敏度测定 PbP2a 的新平台。

A novel platform for high sensitivity determination of PbP2a based on gold nanoparticles composited graphitized mesoporous carbon and doxorubicin loaded hollow gold nanospheres.

机构信息

Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, PR China.

Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, PR China.

出版信息

Biosens Bioelectron. 2016 Mar 15;77:1119-25. doi: 10.1016/j.bios.2015.11.009. Epub 2015 Nov 4.

Abstract

Gold nanoparticles composite graphitized mesoporous carbon nanoparticles (GMCs@AuNPs) biocomposite with the signal amplification capability was successfully synthesized for use in an immunoassay for penicillin binding protein 2 a (PbP2a). The polyamidoamine (PAMAM) dendrimers were first electrodeposited onto the Au electrode can greatly increase the amount of the captured antibodies. Protein A was used to properly orientate immobilized antibody against PbP2a, which strongly improved specificity of the antigen-antibody binding. Hollow gold nanospheres (HGNPs) as effective nanocarriers have been synthesized by sacrificial galvanic replacement of cobalt nanoparticles capable of encapsulating doxorubicin (Dox). The obtained HGNPs@Dox bionanocomposite was used for further loading of detection antibody (Ab2) to form the HGNPs@Dox@Ab2 bioconjugate. Then, the differential pulse voltammetric signals related to the concentration of PbP2a for Dox could be detected, and the immunosensor exhibited a detection limit as low as 0.65 pg mL(-1) (at an S/N ratio of 3). The proposed method with an excellent differentiation ability showed high sensitivity and specificity. The morphologies and electrochemistry properties of the composites were investigated by scanning electron microscopy, electrochemical characterization, UV-visible absorption spectroscopy, fluorescence spectrophotometer and Malvern laser particle size analyzer, respectively. In addition, the basic approach described here would be applicable towards developing biodetection assays against other important targets. Moreover, the bioconjugate of HGNPs@Dox is also a promising pattern to delivery Dox in vivo for anticancer therapy.

摘要

成功合成了具有信号放大能力的金纳米粒子复合石墨化介孔碳纳米粒子(GMCs@AuNPs)生物复合材料,用于青霉素结合蛋白 2a(PbP2a)的免疫分析。首先将聚酰胺-胺(PAMAM)树枝状大分子电沉积到 Au 电极上,可以大大增加捕获抗体的数量。蛋白 A 用于正确定向固定针对 PbP2a 的抗体,这大大提高了抗原-抗体结合的特异性。空心金纳米球(HGNPs)作为有效的纳米载体,通过牺牲电置换钴纳米粒子合成,能够包裹阿霉素(Dox)。所得的 HGNPs@Dox 生物纳米复合材料进一步负载检测抗体(Ab2),形成 HGNPs@Dox@Ab2 生物缀合物。然后,可以检测与 Dox 相关的 PbP2a 浓度的差分脉冲伏安信号,该免疫传感器的检测限低至 0.65 pg mL-1(信噪比为 3)。所提出的方法具有出色的区分能力,表现出高灵敏度和特异性。通过扫描电子显微镜、电化学特性、紫外-可见吸收光谱、荧光分光光度计和马尔文激光粒度分析仪分别研究了复合材料的形貌和电化学性质。此外,这里描述的基本方法可适用于开发针对其他重要靶标的生物检测分析。此外,HGNPs@Dox 的生物缀合物也是体内输送 Dox 用于抗癌治疗的有前途的模式。

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