Graphene oxide-gold nanoparticle-aptamer complexed probe for detecting amyloid beta oligomer by ELISA-based immunoassay

Zhao J 1 , Chang W 2 , Liu L 3 , Xing X 4 , Zhang C 4 , Meng H 4 Show all authors , Gopinath SCB 5 , Lakshmipriya T 6 , Chen Y 7 , Liu Y 8

Affiliations 

  • 1 Department of First Neurology, Baoding No.1 Central hospital, Baoding, Hebei 071000, China. Electronic address: zhaojing1981614@sina.com
  • 2 Endocrine laboratory, Baoding No.1 Central hospital, Baoding, Hebei 071000, China
  • 3 Department of Clinical Psychology, Baoding No.1 Central hospital, Baoding, Hebei 071000, China
  • 4 Department of First Neurology, Baoding No.1 Central hospital, Baoding, Hebei 071000, China
  • 5 Faculty of Chemical Engineering Technology, Universiti Malaysia Perlis (UniMAP), Perlis, Malaysia; Institute of Nano Electronic Engineering, 01000 Kangar, Universiti Malaysia Perlis (UniMAP), Perlis, Malaysia
  • 6 Institute of Nano Electronic Engineering, 01000 Kangar, Universiti Malaysia Perlis (UniMAP), Perlis, Malaysia
  • 7 Department of Oral & Craniofacial Sciences, Faculty of Dentistry, University of Malaya, 50603 Kuala Lumpur, Malaysia
  • 8 Department of First Neurology, Baoding No.1 Central hospital, Baoding, Hebei 071000, China. Electronic address: lyg5976908@sina.com
J Immunol Methods, 2021 02;489:112942.
PMID: 33333060 DOI: 10.1016/j.jim.2020.112942

Abstract

Highly sensitive and easy detection method for Alzheimer's disease (AD) with a suitable biomarker is mandatory for preventing the factors resulting from AD. This research reports a modified ELISA with graphene for the detection of AD biomarker amyloid beta (Aβ) oligomer. Gold nanoparticle (AuNP) conjugated aptamer was used as the capture probe and attached on ELISA-graphene oxide surface through the amine linker. Antibody was used as the detection molecule to reach the maximum detection of Aβ oligomer. Suitable level of APTMS (2%), size of AuNP (30 nm) and aptamer concentration (2 μM) were optimized. This sandwich pattern of aptamer-Aβ oligomer-antibody helps to reach the detection at 50 pM on the optimized ELISA surface and the control experiments in the absence of Aβ oligomer or anti-Aβ oligomer antibody did not show the significant optical detection at 492 nm, indicting the specific detection. Further, Aβ oligomer spiked artificial cerebrospinal fluid did not interfere the detection of Aβ oligomer, confirming the selective detection. This new and modified ELISA surface helps to reach the lower detection of Aβ oligomer and diagnose AD.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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