An amperometric immunosensor based on Nafion-modified electrode for the determination of Schistosoma japonicum antibody

Abstract An amperometric immunosensor is described for the assay of antibody of Schistosoma japonicum (SjAb). The polyanionic perfluorosulfonated Nafion polymer was used to modify the glassy carbon electrode as a platform for the immobilization of S. japonicum antigen (SjAg). The competitive immunoassay format was employed to detect SjAb with horseradish peroxidase (HRP)–SjAb as a tracer, o-aminophenol (o-AP) and hydrogen peroxide as enzyme substrates. The amount of HRP-labeled SjAb binding onto the electrode surface was detected by measuring the product of the enzyme catalytic reaction at −250 mV (versus Ag/AgCl reference electrode). The system was optimized with regard to parameters involved in the preparation of immunosensor and the assay process. The present immunosensor permits a reliable determination of SjAb in the range of 0.05–4.8 μg/ml with a relative standard deviation of below 7.4%. The rabbit serum samples of different infection degree were measured, which implied the immunosensor offer a promising alternative for clinical diagnosis of S. japonicum.

[1]  F. Spener,et al.  An immunosensor based on disposable electrodes for rapid estimation of fatty acid-binding protein, an early marker of myocardial infarction. , 1997, Biosensors & bioelectronics.

[2]  C P Price,et al.  Immunosensors: technology and opportunities in laboratory medicine. , 1996, Clinical chemistry.

[3]  S K Bhatia,et al.  Antibody immobilization using heterobifunctional crosslinkers. , 1997, Biosensors & bioelectronics.

[4]  G. Shen,et al.  A novel approach of antibody immobilization based on n-butyl amine plasma-polymerized films for immunosensors , 2000 .

[5]  H. Budnikov,et al.  Amperometric enzyme immunosensor for the determination of the antigen of the pathogenic fungi Trichophyton rubrum , 2000 .

[6]  G. Shen,et al.  A direct immunoassay for schistosoma japonium antibody (SjAb) in serum by piezoelectric body acoustic wave sensor , 1999 .

[7]  E. Fábregas,et al.  Determination of β-HCG using amperometric immunosensors based on a conducting immunocomposite , 1999 .

[8]  G. Wallace,et al.  Pulsed amperometric detection of thaumatin using antibody-containing poly(pyrrole) electrodes , 1994 .

[9]  J. Zink,et al.  Sol−Gel Encapsulated Anti-Trinitrotoluene Antibodies in Immunoassays for TNT , 2000 .

[10]  Petr Skládal,et al.  Advances in electrochemical immunosensors , 1997 .

[11]  G. Wallace,et al.  Development of a polypyrrole-based human serum albumin sensor , 1991 .

[12]  F. Caruso,et al.  2. Assembly of Alternating Polyelectrolyte and Protein Multilayer Films for Immunosensing , 1997 .

[13]  G. Guilbault,et al.  Amperometric immunosensor for lactate dehydrogenase LD-1. , 1998, Biosensors & bioelectronics.

[14]  P. Thiébaud,et al.  Towards amperometric immunosensor devices. , 1997, Biosensors & bioelectronics.

[15]  H. You,et al.  Covalent coupling of immunoglobulin G to self-assembled monolayers as a method for immobilizing the interfacial-recognition layer of a surface plasmon resonance immunosensor. , 1998, Biosensors & bioelectronics.

[16]  D. Ivnitski,et al.  Portable Immunosensor for the Fast Amperometric Detection of Anti‐Hantavirus Antibodies , 2000 .

[17]  P Atanasov,et al.  Immunosensors: electrochemical sensing and other engineering approaches. , 1998, Biosensors & bioelectronics.

[18]  D. Chan,et al.  Immunosensors--principles and applications to clinical chemistry. , 2001, Clinica chimica acta; international journal of clinical chemistry.

[19]  K R Rogers,et al.  Sol-gel-derived thick-film amperometric immunosensors. , 1998, Analytical chemistry.

[20]  A. M. Smith,et al.  Nature of immobilized antibody layers linked to thioctic acid treated gold surfaces. , 2000, Biosensors & bioelectronics.

[21]  George G. Guilbault,et al.  Amperometric immunosensor for granulocyte-macrophage colony-stimulating factor using screen-printed electrodes , 1999 .

[22]  R A Williams,et al.  Covalent immobilization of protein monolayers for biosensor applications. , 1994, Biosensors & bioelectronics.

[23]  M. Porter,et al.  Immunosensing platforms using spontaneously adsorbed antibody fragments on gold. , 2000, Analytical chemistry.

[24]  G. Shen,et al.  Renewable amperometric immunosensor for Schistosoma japonium antibody assay. , 2001, Analytical chemistry.

[25]  A. Karyakin,et al.  Oriented immobilization of antibodies onto the gold surfaces via their native thiol groups. , 2000, Analytical chemistry.

[26]  G. Radu,et al.  The Construction of an Amperometric Immunosensor for the Thyroid Hormone (+)-3,3′,5-Triiodo-L-Thyronine (L-T3) , 1999 .

[27]  S. Alegret,et al.  Amperometric immunosensors based on rigid conducting immunocomposites. , 1997, Analytical chemistry.