title: Enhanced Fluorescence ELISA Based on HAT Triggering Fluorescence "Turn-on" with Enzyme-Antibody Dual Labeled AuNP Probes for Ultrasensitive Detection of AFP and HBsAg.
abstract: At present, enzyme-linked immunosorbent assay (ELISA) is considered to be the most appropriate approach in clinical biomarker detection, with good specificity, low cost, and straightforward readout. However, unsatisfactory sensitivity severely hampers its wide application in clinical diagnosis. Herein, we designed a new kind of enhanced fluorescence enzyme-linked immunosorbent assay (FELISA) based on the human alpha-thrombin (HAT) triggering fluorescence "turn-on" signals. In this system, detection antibodies (Ab2) and HAT were labeled on the gold nanoparticles (AuNPs) to form the detection probes, and a bisamide derivative of Rhodamine110with fluorescence quenched served as the substrate of HAT. After the sandwich immunoreaction, HAT on the sandwich structure could catalyze the cleavage of the fluorescence-quenched substrate, leading to a strong fluorescence signal for sensing ultralow levels of alpha fetoprotein (AFP) and hepatitis B virus surface antigen (HBsAg). Under the optimized reaction conditions, AFP and HBsAg were detected at the ultralow concentrations of 10-8ng mL-1and 5 × 10-4IU mL-1, respectively, which were at least 104times lower than those of the conventional fluorescence assay and 106times lower than those of the conventional ELISA. In addition, we further discussed the efficiency of the sensitive FELISA in clinical serum samples, showing great potential in practical applications.
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Yudong
authors.initials: Y
authors.lastname: Wu
authors.affiliation: CAS Key Laboratory for Biological Effects of Nanomaterials & Nanosafety National Center for Nanoscience and Technology , Beijing 100190, China.
authors.forename: Weisheng
authors.initials: W
authors.lastname: Guo
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Weipan
authors.initials: W
authors.lastname: Peng
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Jiafang
authors.initials: J
authors.lastname: Piao
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Wu
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Hanjie
authors.initials: H
authors.lastname: Wang
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Xiaoqun
authors.identifier: http://orcid.org/0000-0002-0481-9756
authors.initials: X
authors.lastname: Gong
authors.affiliation: School of Materials Science and Engineering, School of Life Sciences, Tianjin University and Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology (Tianjin) , 92 Weijin Road, Nankai District, Tianjin 300072, P. R. China.
authors.forename: Jin
authors.identifier: http://orcid.org/0000-0002-6752-8526
authors.initials: J
authors.lastname: Chang
mesh_terms: D015374:Biosensing Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D006046:Gold; D006514:Hepatitis B Surface Antigens; D006801:Humans; D053768:Metal Nanoparticles; D013917:Thrombin; D000509:alpha-Fetoproteins
chemical_list: D006514:Hepatitis B Surface Antigens; D000509:alpha-Fetoproteins; D006046:Gold; D013917:Thrombin
keywords: alpha fetoprotein; fluorescence enzyme-linked immunosorbent assay; gold nanoparticles; hepatitis B virus surface antigen; human alpha-thrombin