An electrochemical peptide sensor for detection of dengue fever biomarker NS1.

Dengue virus type 2 NS1 (DENV2 NS1) is a specific and sensitive protein biomarker for dengue fever diagnosis. In this study we used polyvalent phage display to identify unique affinity peptides that can bind NS1 protein. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to investigate the binding interactions. The potential affinity peptide-displayed phage from these methods was selected; its sequence was EHDRMHAYYLTR (R3#10). Amino acid sequence analysis showed that the peptide was rich in basic residues (two His and Arg). Among all the peptides tested, R3#10 showed the greatest decrease in current in CV and increase in impedance in EIS upon binding to NS1 proteins. EIS revealed that R3#10 phage clones were more specific towards NS1 proteins, as compared to bovine serum albumin or the M13 wild type used as control. Detection of NS1 proteins is in accordance with the electron-transfer resistance (Rct) value of the sensor layer, which is confirmed by EIS, and the Kd value of the R3#10 peptide while binding to the phage particles was measured. To the best of our knowledge, this is the first example of identification and characterization of NS1 binding affinity peptides using phage display technology and electrochemical methods. We concluded that these new peptide-displayed phages or free peptides from phages may have potential applications in dengue diagnosis.

[1]  Tuan Vo-Dinh,et al.  DNA bioassay-on-chip using SERS detection for dengue diagnosis. , 2014, The Analyst.

[2]  Li Zhang,et al.  Silicon nanowire biosensor for highly sensitive and rapid detection of Dengue virus , 2010 .

[3]  Lukasz Kurgan,et al.  Unstructural biology of the dengue virus proteins , 2015, The FEBS journal.

[4]  Laura D. Kramer,et al.  Typing of Dengue Viruses in Clinical Specimens and Mosquitoes by Single-Tube Multiplex Reverse Transcriptase PCR , 1998, Journal of Clinical Microbiology.

[5]  Daniel Olson,et al.  Rapid antigen tests for dengue virus serotypes and Zika virus in patient serum , 2017, Science Translational Medicine.

[6]  H. Hwang,et al.  Selection of High Affinity Peptides for Prediction of Colorectal Adenoma–to‐Carcinoma Progression , 2016 .

[7]  Tae Jung Park,et al.  High sensitive and selective electrochemical biosensor: Label-free detection of human norovirus using affinity peptide as molecular binder. , 2017, Biosensors & bioelectronics.

[8]  Jong Pil Park,et al.  Evolutionary identification of affinity peptides for the detection of sepsis biomarker procalcitonin , 2015 .

[9]  V. L. Carvalho,et al.  A Luminex-based single DNA fragment amplification assay as a practical tool for detecting and serotyping dengue virus. , 2016, Journal of virological methods.

[10]  D. Tai,et al.  Development of immunochips for the detection of dengue viral antigens , 2003 .

[11]  Oleksiy Krupin,et al.  Serological diagnosis of dengue infection in blood plasma using long-range surface plasmon waveguides. , 2014, Analytical chemistry.

[12]  A. Mulchandani,et al.  A heparin-functionalized carbon nanotube-based affinity biosensor for dengue virus. , 2017, Biosensors & bioelectronics.

[13]  W. Coltro,et al.  Dengue diagnosis on laser printed microzones using smartphone-based detection and multivariate image analysis , 2016 .

[14]  M. Tabrizian,et al.  Microfluidic designs and techniques using lab-on-a-chip devices for pathogen detection for point-of-care diagnostics. , 2012, Lab on a chip.

[15]  O. Parkash,et al.  Screen Printed Carbon Electrode Based Electrochemical Immunosensor for the Detection of Dengue NS1 Antigen , 2014, Diagnostics.

[16]  Donald M Cropek,et al.  High affinity peptides for the recognition of the heart disease biomarker troponin I identified using phage display , 2009, Biotechnology and bioengineering.

[17]  L. Kubota,et al.  Electrochemical detection of dengue virus NS1 protein with a poly(allylamine)/carbon nanotube layered immunoelectrode , 2015 .

[18]  Donald M. Cropek,et al.  Rapid Development of New Protein Biosensors Utilizing Peptides Obtained via Phage Display , 2011, PloS one.

[19]  S. Sidhu,et al.  Engineering M13 for phage display. , 2001, Biomolecular engineering.

[20]  H. Hwang,et al.  Identification of high affinity peptides for capturing norovirus capsid proteins , 2015 .

[21]  Kimberly Hamad-Schifferli,et al.  Multicolored silver nanoparticles for multiplexed disease diagnostics: distinguishing dengue, yellow fever, and Ebola viruses. , 2015, Lab on a chip.

[22]  A. R. Ruslinda,et al.  Electrical detection of dengue virus (DENV) DNA oligomer using silicon nanowire biosensor with novel molecular gate control. , 2016, Biosensors & bioelectronics.

[23]  Zhiqiang Gao,et al.  Quantum dots and duplex-specific nuclease enabled ultrasensitive detection and serotyping of Dengue viruses in one step in a single tube. , 2015, Biosensors & bioelectronics.

[24]  J. d'Alayer,et al.  Secreted dengue virus nonstructural protein NS1 is an atypical barrel-shaped high-density lipoprotein , 2011, Proceedings of the National Academy of Sciences.

[25]  S. Sankar,et al.  Development and multi-use applications of dengue NS1 monoclonal antibody for early diagnosis , 2017 .

[26]  Richard A Montagna,et al.  Microfluidic biosensor for the serotype-specific detection of dengue virus RNA. , 2005, Analytical chemistry.

[27]  Hsien-Chang Chang,et al.  A bead-based immunofluorescence-assay on a microfluidic dielectrophoresis platform for rapid dengue virus detection. , 2017, Biosensors & bioelectronics.

[28]  Anja Boisen,et al.  Quantification of NS1 dengue biomarker in serum via optomagnetic nanocluster detection , 2015, Scientific Reports.

[29]  Kanti Pabbaraju,et al.  Simultaneous detection of Zika, Chikungunya and Dengue viruses by a multiplex real-time RT-PCR assay. , 2016, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[30]  F. Ibrahim,et al.  Synthesis and characterization of methacrylic microspheres for biomolecular recognition: Ultrasensitive biosensor for Dengue virus detection , 2014 .

[31]  Stephen J. Thomas.  Developing a dengue vaccine: progress and future challenges , 2014, Annals of the New York Academy of Sciences.

[32]  C. P. de Melo,et al.  An impedimetric biosensor for detection of dengue serotype at picomolar concentration based on gold nanoparticles-polyaniline hybrid composites. , 2011, Colloids and surfaces. B, Biointerfaces.