Optical Immunosensors for the Efficient Detection of Target Biomolecules

이도현,황장선,서영민,Assaf A. Gilad,최종훈 한국생물공학회 2018 Biotechnology and Bioprocess Engineering Vol.23 No.2

Recently, immunosensors have attracted attention because they are widely applied for the detection of various pathogens. Among the commonly used immunosensors, the optical immunosensor features prominently as an effective tool for the quantification of the amount of antibodies, antigens, or haptens in complex samples with high sensitivity and specificity. However, very few studies provide comprehensive overviews of optical immunosensors. In this review, we present various methods and applications of optical immunosensors in pathogen detection. We introduced a concise definition of optical immunosensors and the principle of using them for detection. We subsequently discuss the main categories of optical immunosensors and their application to the detection of pathogens, as well as their advantages and limitations. Recent publications from 2006 to 2015 on variously designed optical immunosensors have also been updated. We conclude the review with a brief summary and discuss future directions of optical immunosensors.

Dual-responsive immunosensor that combines colorimetric recognition and electrochemical response for ultrasensitive detection of cancer biomarkers

Hong, Wooyoung,Lee, Sooyeon,Cho, Youngnam Elsevier 2016 Biosensors & bioelectronics Vol.86 No.-

<P><B>Abstract</B></P> <P>We developed a nanoroughened, biotin-doped polypyrrole immunosensor for the detection of tumor markers through dual-signal (electrochemical and colorimetric) channels, electrochemical and colorimetric, that demonstrates remarkable analytical performance. A rapid, one-step electric field-mediated method was employed to fabricate the immunosensor with nanoscale roughness by simply modulating the applied electrical potential. We demonstrated the successful detection of three tumor markers (CA125, CEA, and PSA) via the double enzymatic signal amplifications in the presence of a target antigen, ultimately leading to desired diagnostic accuracy and reliability. The addition of multiple horseradish peroxidase (HRP)- and antibody-labeled nanoparticles greatly amplified the signal and simplified the measurement of cancer biomarker proteins by sequentially magnifying electrochemical and colorimetric signals in a single platform. The two parallel assays performed using the proposed immunosensor have yielded highly consistent and reproducible results. Additionally, for the analysis of plasma samples in a clinical setting, the values obtained with our immunosensor were validated by correlating the results with those of a standard radioimmunoassay (RIA), which obtained very similar clinically valid responses.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A rapid, one-step electric field-mediated method was developed to fabricate an immunosensor. </LI> <LI> The performance of immunosensor was significantly increased with increasing applied potential. </LI> <LI> The immunosensor measures subtle variations in electrochemical and colorimetric signals. </LI> <LI> The two parallel assays performed on a single platform have yielded highly consistent results. </LI> </UL> </P>

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 Elsevier 2017 Analytica chimica acta Vol.967 No.-

<P><B>Abstract</B></P> <P>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.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A novel fusion protein was developed as crosslinker for rapid self-oriented immobilization of antibody on SiNPs. </LI> <LI> This crosslinker contributed to easy formation of SiNPs/Ab complexes, resulting in electrochemical signal enhancement. </LI> <LI> Novel prevention method of electrode fouling was developed to enhance sensitivity of the electrochemical immunosensor. </LI> <LI> This immunosensor exhibited excellent sensitivity with the simple fabrication methods. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Nonenzymatic Electrochemical Immunosensor Using Ferroferric Oxide–Manganese Dioxide–Reduced Graphene Oxide Nanocomposite as Label for α-Fetoprotein Detection

Wensi Jian,Chunping Wang,Zuanguang Chen,Yanyan Yu,Duanping Sun,Liping Han,Lijuan Shi 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2016 NANO Vol.11 No.10

A novel nonenzymatic electrochemical immunosensor was fabricated for quantitative detection of α-fetoprotein (AFP). The immunosensor was constructed by modifying gold electrode with electrochemical reduction of graphene oxide-carboxyl multi-walled carbon nanotube composites (ERGO–CMWCNTs) and electrodeposition of gold nanoparticles (AuNPs) for effective immobilization of primary antibody (Ab1). Ferroferric oxide–manganese dioxide–reduced graphene oxide nanocomposites (Fe3O4@MnO2–rGO) were designed as labels for signal amplification. On one hand, the excellent electroconductivity and outstanding electron transfer capability of ERGO–CMWCNTs/AuNPs improved the sensitivity of the immunosensor. On the other hand, introduction of rGO could not only increase the specific surface area for immobilization of secondary antibody (Ab2) but also build a synergetic effect to reinforce the electrocatalytic properties of catalysts. Fe3O4@MnO2–rGO nanocomposites were characterized by scanning electron microscope, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Using AFP as a model analyte, the proposed sandwich-type electrochemical immunosensor exhibited a wide linear range of 0.01–50 ng · mL-1 with a low detection limit of 5.8 pg · mL-1. Moreover, the Fe3O4@MnO2–rGO-based peroxidase mimetic system displayed an excellent analytical performance with low cost, satisfactory reproducibility and high selectivity, which could be further extended for detecting other disease-related biomarkers.

Cell surface-camouflaged graphene oxide immunosensor for identifying immune reactions

Hwang, Yong Hwa,Jeon, Eun Ae,Lee, Dong Yun Elsevier 2018 Journal of industrial and engineering chemistry Vol.59 No.-

<P><B>Abstract</B></P> <P>Monitoring the intensity of immune reactions is very useful to diagnose the fate of transplanted cells. For directly monitoring immune reactions after pancreatic islet cell transplantation, here graphene oxide (GO) quencher was chemically conjugated with fluorescence dye via Granzyme B (GrB)-specific peptide, so called to GO-based immunosensor. GO-based immunosensor was stably immobilized onto surface of living cells without damage of viability and functionality. The cell-surface camouflaged GO-based immunosensor could detect GrB (>2unit/ml) secreted from immune cells, thereby dequenching fluorescence. Collectively, we anticipated that the GO-based immunosensor could be utilized to monitor cell graft rejection after cell therapy <I>in vivo</I>.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Cell surface-camouflaged graphene oxide immunosensor for identifying immune reactions

황용화,전은애,이동윤 한국공업화학회 2018 Journal of Industrial and Engineering Chemistry Vol.59 No.-

Monitoring the intensity of immune reactions is very useful to diagnose the fate of transplanted cells. For directly monitoring immune reactions after pancreatic islet cell transplantation, here graphene oxide (GO) quencher was chemically conjugated with fluorescence dye via Granzyme B (GrB)-specific peptide, so called to GO-based immunosensor. GO-based immunosensor was stably immobilized onto surface of living cells without damage of viability and functionality. The cell-surface camouflaged GO-based immunosensor could detect GrB (>2 unit/ml) secreted from immune cells, thereby dequenching fluorescence. Collectively, we anticipated that the GO-based immunosensor could be utilized to monitor cell graft rejection after cell therapy in vivo.

Development of a paper-based electrochemical immunosensor using an antibody-single walled carbon nanotubes bio-conjugate modified electrode for label-free detection of foodborne pathogens

Bhardwaj, Jyoti,Devarakonda, Sivaranjani,Kumar, Suveen,Jang, Jaesung Elsevier Sequoia 2017 Sensors and actuators. B Chemical Vol.253 No.-

<P><B>Abstract</B></P> <P>The need for low-cost, sensitive, and reliable sensors for the detection of whole bacterial cells in food samples without pre-treatment has been increasing. Outbreaks of foodborne diseases can be severe, especially in developing countries; however, most bio-detection tools are unaffordable. Here, we have developed a rapid and low-cost paper-based electrochemical immunosensor for label-free detection of <I>Staphylococcus aureus</I>, using antibody (Ab)-single walled carbon nanotube (SWCNT) bio-conjugates. Anti-<I>S. aureus</I> antibodies were covalently attached onto the SWCNTs, using the <I>N</I>-(3-dimethylaminopropyl)-<I>N</I>’-ethylcarbodiimide hydrochloride/<I>N</I>-hydroxysuccinimide coupling reagent. These Ab-SWCNT bio-conjugates were then immobilized on the working electrode, and the presence of <I>S. aureus</I> was detected by analyzing the change in peak current following antigen-antibody complex formation. Differential pulse voltammetry was performed with a bacterial concentration ranging from 10 to 10<SUP>7</SUP> colony forming units (CFU) mL<SUP>−1</SUP>. A selectivity assay using <I>Escherichia coli</I> B<I>, Bacillus subtilis</I>, and <I>S. epidermidis</I> (to examine cross-reactivity) showed that the sensor was specific to <I>S. aureus.</I> Moreover, this immunosensor showed a good linear relationship (R<SUP>2</SUP> =0.976) between the increase in peak current and logarithmic <I>S. aureus</I> concentration, with a rapid detection time (30min) and a limit of detection of 13CFUmL<SUP>−1</SUP> in spiked milk samples. This low-cost immunosensor can be used for rapid detection of pathogens in actual food samples with high sensitivity and specificity.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A sensitive paper based electrochemical sensor for label-free detection of bacteria in food samples is presented. </LI> <LI> Ab-SWCNT conjugate was used to decrease the functionalization steps and increase the robustness of the immunosensors. </LI> <LI> A detection limit of 13 CFUmL-1 in spiked milk samples was obtained with rapid detection time (∼30min). </LI> <LI> A highly linear behavior and high specificity were observed. </LI> </UL> </P>

Ultrasensitive dual probe immunosensor for the monitoring of nicotine induced-brain derived neurotrophic factor released from cancer cells

Akhtar, Mahmood H.,Hussain, Khalil K.,Gurudatt, N.G.,Chandra, Pranjal,Shim, Yoon-Bo Elsevier 2018 Biosensors & bioelectronics Vol.116 No.-

<P><B>Abstract</B></P> <P>Brain-derived neurotrophic factor (BDNF) was detected in the extracellular matrix of neuronal cells using a dual probe immunosensor (DPI), where one of them was used as a working and another bioconjugate loading probe. The working probe was fabricated by covalently immobilizing capture anti-BDNF (Cap Ab) on the gold nanoparticles (AuNPs)/conducting polymer composite layer. The bioconjugate probe was modified by drop casting a bioconjugate particles composed of conducting polymer self-assembled AuNPs, immobilized with detection anti-BDNF (Det Ab) and toluidine blue O (TBO). Each sensor layer was characterized using the surface analysis and electrochemical methods. Two modified probes were precisely faced each other to form a microfluidic channel structure and the gap between inside modified surfaces was about 19 µm. At optimized conditions, the DPI showed a linear dynamic range from 4.0 to 600.0 pg/ml with a detection limit of 1.5 ± 0.012 pg/ml. Interference effect of IgG, arginine, glutamine, serine, albumin, and fibrinogene were examined and stability of the developed biosensor was also investigated. The reliability of the DPI sensor was evaluated by monitoring the extracellular release of BDNF using exogenic activators (ethanol, K<SUP>+</SUP>, and nicotine) in neuronal and non-neuronal cells. In addition, the effect of nicotine onto neuroblastoma cancer cells (SH-SY5Y) was studied in detail.</P> <P><B>Highlights</B></P> <P> <UL> <LI> This work reports for the first time a dual probe immunosensor for BDNF. </LI> <LI> Bioconjugate particles were synthesized, which composed of anti-BDNF and TBO chemically bonded onto the SAM. </LI> <LI> The proposed method can be extended for <I>in-vivo</I> applications, which is a unique feature of this immunosensor. </LI> <LI> In biological matrix, nicotine-induced BDNF release in neuroblastoma cancer cells was monitored. </LI> </UL> </P>

Gold nanoparticle-based signal augmentation of quartz crystal microbalance immunosensor measuring C-reactive protein

Kim, N.,Kim, D.K.,Cho, Y.J. Elsevier 2010 CURRENT APPLIED PHYSICS Vol.10 No.4

A biotinylated anti-rat C-reactive protein (CRP) antibody was ingeniously prepared by the reaction of the unmodified antibody with a water-soluble sulfosuccinimidyl-6-(biotinamido)hexanoate. The molar biotin incorporation of the resulting modified antibody was found as 5.82. A flow-type indirect-competitive quartz crystal microbalance immunosensor system was constructed with the sensor chip immobilized with 2mg/mL CRP as the coating antigen. When 200μL of the modified antibody having the concentration of 0.250mg/mL was added with a streptavidin-coated gold nanoparticle (GNP) to the immunosensor system, the frequency shift obtained was 139.8+/-0.3Hz. Compared to the frequency shift of 91.1+/-1.3Hz found with the addition of the unmodified antibody only, the signal augmentation after GNP binding amounted to 53.4%, which resulted in sensitivity improvement of the current immunosensor.

Gold nanoparticle-based signal augmentation of quartz crystal microbalance immunosensor measuring C-reactive protein

김남수,D.-K. Kim,Y.-J. Cho 한국물리학회 2010 Current Applied Physics Vol.10 No.4

A biotinylated anti-rat C-reactive protein (CRP) antibody was ingeniously prepared by the reaction of the unmodified antibody with a water-soluble sulfosuccinimidyl-6-(biotinamido)hexanoate. The molar biotin incorporation of the resulting modified antibody was found as 5.82. A flow-type indirect-competitive quartz crystal microbalance immunosensor system was constructed with the sensor chip immobilized with 2 mg/mL CRP as the coating antigen. When 200 μL of the modified antibody having the concentration of 0.250 mg/mL was added with a streptavidin-coated gold nanoparticle (GNP) to the immunosensor system, the frequency shift obtained was 139.8 ± 0.3 Hz. Compared to the frequency shift of 91.1 ± 1.3 Hz found with the addition of the unmodified antibody only, the signal augmentation after GNP binding amounted to 53.4%, which resulted in sensitivity improvement of the current immunosensor.