title: Simultaneous electrochemical immunoassay using graphene-Au grafted recombinant apoferritin-encoded metallic labels as signal tags and dual-template magnetic molecular imprinted polymer as capture probes.
abstract: A novel electrochemical multiplexed immunoassay was designed for simultaneous determination of alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) using recombinant apoferritin-encoded metallic nanoparticles (rApo-M) as labels and dual-template magnetic molecularly imprinted polymers (MMIPs) as capture probes. The labels were prepared by loading recombinant apoferritin (r-Apo) and separately immobilize primary antibodies (anti-AFP and anti-CEA) via Au nanoparticles of in site growth on graphene (G). The capture probes were synthesized by self-polymerization of dopamine (DA) on the Fe3O4 nanoparticles (Fe3O4 NPs) and using AFP and CEA as the template proteins, which were used to enrich the targets simultaneously. After a sandwich-type immunoreaction, the labels were captured to the surface of MMIPs. The subsequent electrochemical stripping analysis of the metal components from the immunocomplex provide a means for quantification of targets based on the peak currents of Cd and Pb. Experimental results showed the immunoassay enabled the simultaneous determination of AFP and CEA in a single run with wide dynamic ranges of 0.001-5ngmL(-1). And the detection limits of AFP and CEA were 0.3 and 0.35pgmL(-1) (S/N=3), respectively. These results suggested that the proposed multiplexed immunoassay would be applied for clinical screening of other biomarkers.
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China.
authors.forename: De
authors.initials: D
authors.lastname: Wang
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China. Electronic address: ganning@nbu.edu.cn.
authors.forename: Ning
authors.initials: N
authors.lastname: Gan
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China.
authors.forename: Huairong
authors.initials: H
authors.lastname: Zhang
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China.
authors.forename: Tianhua
authors.initials: T
authors.lastname: Li
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China.
authors.forename: Li
authors.initials: L
authors.lastname: Qiao
authors.affiliation: State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, PR China.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Faculty of Marine, Ningbo University, Ningbo 315211, PR China.
authors.forename: Xiurong
authors.initials: X
authors.lastname: Su
authors.affiliation: The Department of Orthopaedics and Traumatology, Southern Medical University, Guangzhou, Guangdong 510515, PR China.
authors.forename: Shan
authors.initials: S
authors.lastname: Jiang
mesh_terms: D001052:Apoferritins / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D002272:Carcinoembryonic Antigen / Q000032:analysis / Q000097:blood*; D055664:Electrochemical Techniques / Q000379:methods*; D006046:Gold / Q000737:chemistry; D006108:Graphite / Q000737:chemistry; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D057230:Limit of Detection; D059346:Magnets / Q000737:chemistry*; D054802:Molecular Imprinting; D011108:Polymers / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D000509:alpha-Fetoproteins / Q000032:analysis*
chemical_list: D002272:Carcinoembryonic Antigen; D055767:Immobilized Proteins; D011108:Polymers; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins; D006046:Gold; D006108:Graphite; D001052:Apoferritins
keywords: Dual-template magnetic molecularly imprinted polymers; Graphene–Au; Multiplexed immunoassay; Recombinant apoferritin