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Functional fusion proteins and prevention of electrode fouling for a sensitive electrochemical immunosensor
- Functional fusion proteins and prevention of electrode fouling for a sensitive electrochemical immunosensor
- Kim, A-Ram; Park, Tae Jung; Kim, Minseok S.; Kim, In-Ho; Kim, Ki-Suk; Chung, Kwang Hoe; Ko, Sungho
- DGIST Authors
- Kim, Minseok S.
- Issue Date
- Analytica Chimica Acta, 967, 70-77
- Article Type
- Alkalinity; Antibodies; Bins; Biosensors; Carbon Nanotubes; Chemical Bonds; Conformational Changes; Cyclic Olefin Copolymer; Cyclic Olefin Copolymers; Dendrimer; Diseases; Electro Chemical Electrodes; Electrochemical Immunosensor; Electrochemical Immunosensors; Electrode Fouling; Electrodes; Fouling; Gold; Gold Deposits; Gold Nanoparticles; Horseradish Peroxidase; Immunosensors; Nanoparticles; Olefins; Orientation; Phosphatases; Polypeptides; Prostate Specific Antigen; Protein G; Proteins; Recombinant Proteins; Silica; Silica Binding Polypeptide; Silica Nanoparticle; Silica Nanoparticles; Silica Nanoparticles; Surface Concentration
- A highly sensitive electrochemical immunosensor was developed by preventing electrode fouling and using a novel fusion protein of silica binding polypeptides (SBP)-protein G (ProG) created by recombinant DNA technology as a functional crosslinker for rapid and self-oriented immobilization of antibodies onto silica nanoparticles (SiNPs). Antibody immobilization onto the SiNPs by the SBP-ProG could rapidly be achieved without any chemical treatment. The immunosensor was fabricated through bonding of a partially gold-deposited cyclic olefin copolymer (COC) (top substrate) and gold patterned interdigitated array COC electrode (bottom substrate). To prevent electrode fouling, human immunoglobulin G (hIgG) was immobilized onto the ceiling inside the microchannel, instead of the bottom electrode. Alkaline phosphatase (AP)-labeled anti-hIgG was allowed to immunoreact with hIgG on the ceiling, followed by addition of an enzyme to generate an oxidative peak current. A three-fold increase in current was observed from the immunosensor without any electrode fouling compared with a control with the protein functionalized electrode. Also, the SiNPs facilely coated with AP-anti-hIgG via the SBP-ProG could increase the electrochemical signal up to 20% larger than that of the AP-anti-hIgG alone. Furthermore, this immunosensor was ultrasensitive with a detection limit of 0.68 pg/mL of a biomarker associated with prostate cancer. © 2017 Elsevier B.V.
- Elsevier B.V.
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