A label-free electrochemical immunosensor for hepatitis B based on hyaluronic acid-carbon nanotube hybrid film.

An electrochemical immunosensor developed for detection of antibodies to hepatitis B core protein (anti-HBc) is described. Anti-HBc is the earliest serological marker from hepatitis B virus (HBV) infection, remaining all life after contact with virus, being considered the most important marker for uses in screening of blood bank. A nanohybrid surface assembled onto a glassy carbon electrode consisting of amino carbon nanotubes recovered by hyaluronic acid was used as sensing platform to detect the anti-HBc. All the steps of electrode surface modification were characterized by Scanning Electronic Microscopy and extensively evaluated by electrochemical techniques. The electrode response was measured by direct anti-HBc antigen interactions by square wave voltammetry, dispensing uses of label or chemical mediators. Under optimal conditions, the anodic peak current which was proportional to the anti-HBs concentration. The immunosensor response was linear toward anti-HBc in concentrations up to 6 ng mL(-1), with a detection limit of 0.03 ng mL(-1). The linear range achieved was according to clinical level, indicating the immunosensor as promising tool for use as a criterion for blood bag disposal. The enhancement of the hyaluronic acid by carbon nanotube promoted an increase of charge electron transfer, besides a stable platform for HBc.

[1]  B. Piro,et al.  A label-free electrochemical immunosensor for direct, signal-on and sensitive pesticide detection. , 2012, Biosensors & bioelectronics.

[2]  B. Cowie,et al.  Hepatitis B virus epidemiology. , 2015, Cold Spring Harbor perspectives in medicine.

[3]  Lan Xu,et al.  Electrochemical tolazoline sensor based on gold nanoparticles and imprinted poly-o-aminothiophenol film , 2010 .

[4]  Jian Huang,et al.  Developing a double-antigen sandwich ELISA for effective detection of human hepatitis B core antibody. , 2008, Comparative immunology, microbiology and infectious diseases.

[5]  Bin Du,et al.  Label-free immunosensor for the detection of kanamycin using Ag@Fe₃O₄ nanoparticles and thionine mixed graphene sheet. , 2013, Biosensors & bioelectronics.

[6]  R. F. Dutra,et al.  A carbon nanotube screen-printed electrode for label-free detection of the human cardiac troponin T. , 2013, Talanta.

[7]  M. Moeller,et al.  Tailored hyaluronic acid hydrogels through hydrophilic prepolymer cross-linkers , 2010 .

[8]  A. Lok,et al.  Prevention of posttransfusion hepatitis B and C by screening for antibody to hepatitis C virus and antibody to HBcAg , 1993, Hepatology.

[9]  Amruta S. Patil,et al.  Hepatitis B Diagnosis in Blood Bank: Evaluation and Challenges , 2015 .

[10]  Joseph Wang Carbon‐Nanotube Based Electrochemical Biosensors: A Review , 2005 .

[11]  B. Rehermann,et al.  Immunology of hepatitis B virus and hepatitis C virus infection , 2005, Nature Reviews Immunology.

[12]  Yon Hui Kim,et al.  A nanowire-based label-free immunosensor: direct incorporation of a PSA antibody in electropolymerized polypyrrole. , 2014, Biosensors & bioelectronics.

[13]  Robert Stern,et al.  Hyaluronic acid: a natural biopolymer with a broad range of biomedical and industrial applications , 2006, Biotechnology Letters.

[14]  M. D. Rooij,et al.  Electrochemical Methods: Fundamentals and Applications , 2003 .

[15]  P. Holland,et al.  Significance of antibody to hepatitis B core antigen in blood donors as determined by their serologic response to hepatitis B vaccine , 1993, Transfusion.

[16]  R. F. Dutra,et al.  A carbon nanotube-based electrochemical immunosensor for cardiac troponin T , 2013 .

[17]  M. Emara,et al.  Occult hepatitis B infection in egyptian chronic hepatitis C patients: prevalence, impact on pegylated interferon/ribavirin therapy , 2010, Virology Journal.

[18]  Hedayatollah Ghourchian,et al.  A gold nanoparticle-based immunosensor for the chemiluminescence detection of the hepatitis B surface antigen , 2014 .

[19]  T. Aastrup,et al.  Optimizing immobilization on two-dimensional carboxyl surface: pH dependence of antibody orientation and antigen binding capacity. , 2010, Analytical biochemistry.

[20]  B. McMahon,et al.  Chronic hepatitis B: Update 2009 , 2009, Hepatology.

[21]  Jinghua Yu,et al.  Electrochemical sensor based on molecularly imprinted film at polypyrrole-sulfonated graphene/hyaluronic acid-multiwalled carbon nanotubes modified electrode for determination of tryptamine. , 2012, Biosensors & bioelectronics.

[22]  R. F. Dutra,et al.  A sensor tip based on carbon nanotube-ink printed electrode for the dengue virus NS1 protein. , 2013, Biosensors & bioelectronics.

[23]  Y. Takai,et al.  Hyaluronan-based Biomaterials in Tissue Engineering , 2004 .

[24]  Rahman,et al.  A Cyclic Voltammetric Study of the Redox Reaction of Cu(II) in Presence of Ascorbic Acid in Different pH Media , 2013 .

[25]  P. Poulin,et al.  Carbon nanotubes induced gelation of unmodified hyaluronic acid. , 2013, Langmuir : the ACS journal of surfaces and colloids.

[26]  R. Ellis Hepatitis B Vaccines in Clinical Practice , 1992 .

[27]  D. Lavanchy,et al.  Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures , 2004, Journal of viral hepatitis.

[28]  Wei-ling Fu,et al.  Biosensors for hepatitis B virus detection. , 2014, World journal of gastroenterology.

[29]  Frank Davis,et al.  Recent trends in antibody based sensors. , 2012, Biosensors & bioelectronics.