Aptamer-MIP hybrid receptor for highly sensitive electrochemical detection of prostate specific antigen

被引:224
作者
Jolly, Pawan [1 ]
Tamboli, Vibha [2 ]
Harniman, Robert L. [3 ]
Estrela, Pedro [1 ]
Allender, Chris J. [2 ]
Bowen, Jenna L. [2 ]
机构
[1] Univ Bath, Dept Elect & Elect Engn, Bath BA2 7AY, Avon, England
[2] Cardiff Univ, Sch Pharm & Pharmaceut Sci, Cardiff CF10 3NB, S Glam, Wales
[3] Univ Bristol, Sch Chem, Bristol BS8 1TS, Avon, England
关键词
Molecular imprinting; Aptamer; Electrochemical impedance spectroscopy; Prostate cancer; Prostate specific antigen; MOLECULARLY IMPRINTED POLYMERS; EAU GUIDELINES; DNA APTAMER; BIOSENSOR; CANCER; PROTEINS; SENSORS; CHARGE; FILM;
D O I
10.1016/j.bios.2015.08.043
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
This study reports the design and evaluation of a new synthetic receptor sensor based on the amalgamation of biomolecular recognition elements and molecular imprinting to overcome some of the challenges faced by conventional protein imprinting. A thiolated DNA aptamer with established affinity for prostate specific antigen (PSA) was complexed with PSA prior to being immobilised on the surface of a gold electrode. Controlled electropolymerisation of dopamine around the complex served to both entrap the complex, holding the aptamer in, or near to, it's binding conformation, and to localise the PSA binding sites at the sensor surface. Following removal of PSA, it was proposed that the molecularly imprinted polymer (MIP) cavity would act synergistically with the embedded aptamer to form a hybrid receptor (apta-MIP), displaying recognition properties superior to that of aptamer alone. Electrochemical impedance spectroscopy (EIS) was used to evaluate subsequent rebinding of PSA to the apta-MIP surface. The apta-MIP sensor showed high sensitivity with a linear response from 100 pg/ml to 100 ng/ml of PSA and a limit of detection of 1 pg/ml, which was three-fold higher than aptamer alone sensor for PSA. Furthermore, the sensor demonstrated low cross-reactivity with a homologous protein (human Kallilcrein 2) and low response to human serum albumin (HSA), suggesting possible resilience to the non-specific binding of serum proteins. (C) 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
引用
收藏
页码:188 / 195
页数:8
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