A novel reverse fluorescent immunoassay approach for sensing human chorionic gonadotropin based on silver-gold nano-alloy and magnetic nanoparticles

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• 作者：Xiaopeng Huang ; Yuqin Li ; Xiang Huang ; Xiaona Xie…
• 关键词：Silver ; gold nano ; alloy ; Reverse fluorescent immunoassay ; Human chorionic gonadotropin ; Magnetic nanoparticles
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2016
• 出版时间：January 2016
• 年：2016
• 卷：408
• 期：2
• 页码：619-627
• 全文大小：918 KB
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• 作者单位：Xiaopeng Huang (1)
  Yuqin Li (2)
  Xiang Huang (1)
  Xiaona Xie (1)
  Yanping Xu (3)
  Yaowen Chen (3)
  Wenhua Gao (1) (3)
  
  1. Department of Chemistry and Laboratory for Preparation and Application of Ordered Structural Materials of Guangdong Province, Shantou University, Shantou, Guangdong, 515063, China
  2. Department of Pharmacy, Taishan Medicine College, Taian, Shandong, 271016, China
  3. Analysis & Testing Center, Shantou University, Shantou, Guangdong, 515063, China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Food Science
  Inorganic Chemistry
  Physical Chemistry
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1618-2650

文摘

A novel and environmentally friendly reverse fluorescent immunoassay approach was proposed and utilized for sensing human chorionic gonadotropin (HCG) in human serum by coupling a newly prepared and highly fluorescent glutathione-stabilized silver-gold nano-alloy (GSH-AgAuNAs) with magnetic nanoparticles (MNPs). To construct such a reverse system, fluorescent GSH-AgAuNAs and MNPs were first prepared and bio-functionalized with monoclonal antibodies (Mab-I and Mab-II) toward HCG antigen, respectively. Then, the GSH-AgAuNAs functionalized with Mab-I were incubated with HCG, followed by the addition of MNPs attached to Mab-II. Thereafter, a sandwich-type immunoassay could be constructed for determination of HCG owing to the antibody-antigen recognition between the functionalized GSH-AgAuNAs and MNPs. Afterwards, a magnetic collection was employed. Hence, the amount of GSH-AgAuNAs would be reduced through an immuno-magnetic separation, thus weakening the fluorescent intensity. Different from conventional immunoassay, our work determined the quantitative signal by measuring the decreasing gradient fluorescent intensity. Under optimal conditions, the developed reverse method exhibited a wide linear range of 0.5–600 ng mL−1 toward HCG with a detection limit of 0.25 ng mL−1. Additionally, the proposed immunoassay was validated using spiked samples, illustrating a satisfactory result in practical application. Keywords Silver-gold nano-alloy Reverse fluorescent immunoassay Human chorionic gonadotropin Magnetic nanoparticles