A model of optical immunosensor for hemoproteins based on Langmuir-Blodgett films of FITC-labelled immunoglobulin G

Abstract A direct optical method based on the interaction of hemoproteins with a thin film modified with a fluorescent label specific immunoglobulin G (IgG) has been developed for the fluorimetric determination of antigen–antibody binding. The sensing surface is composed of a Langmuir–Blodgett (LB) film of fluorescein isothiocyanate (FITC)-modified IgG. The LB film binds heme-containing antigen and changes its fluorescent properties owing to resonant energy transfer in the donor–acceptor pair FITC–heme. The quenching of FITC fluorescence emission grows under increasing concentrations of hemoproteins over a broad range; the quenching degree depends on the molecular weight of the hemoprotein. The main advantage of the proposed method is the possibility of developing a direct immunosensor based on evanescent excited fibre optics.

[1]  B. Bluestein,et al.  Fiber optic evanescent wave immunosensors for medical diagnostics. , 1990, Trends in biotechnology.

[2]  J. Herron,et al.  Characterization of immobilized antibodies on silica surfaces , 1988, IEEE Transactions on Biomedical Engineering.

[3]  J. Andrade,et al.  The influence of adsorption of native and modified antibodies on their activity. , 1989, Journal of immunological methods.

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

[5]  Tuan Vo-Dinh,et al.  Immunosensors: Principles and Applications , 1993 .

[6]  U. Jönsson,et al.  Immobilization of immunoglobulins on silica surfaces. Stability. , 1985, The Biochemical journal.

[7]  R. M. Sutherland,et al.  Opto-electronic immunosensors: a review of optical immunoassay at continuous surfaces. , 1985, Biosensors.

[8]  R Renneberg,et al.  Surface investigations on the development of a direct optical immunosensor. , 1998, Biosensors & bioelectronics.

[9]  I. Turko,et al.  Stability and stabilization of immunoglobulin G Langmuir-Blodgett films , 1993 .

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

[11]  M. Lösche,et al.  Novel biosensoric devices based on molecular protein hetero-multilayer films. , 1997, Advances in biophysics.

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

[13]  M. Aizawa,et al.  Preparation and characterization of protein Langmuir-Blodgett films , 1989 .

[14]  J. Lakowicz Principles of fluorescence spectroscopy , 1983 .

[15]  C. Nicolini,et al.  Immunological activity of IgG Langmuir films oriented by protein A sublayer , 1995 .

[16]  D. Reeder,et al.  Anti-T2 Monoclonal Antibody Immobilization On Quartz Fibers: Stability and Recognition of T2 Mycotoxin , 1989 .

[17]  R. Poljak,et al.  Three-dimensional structure of immunoglobulins. , 1979, Annual review of biochemistry.

[18]  I. Turko,et al.  Langmuir-Blodgett films of immunoglobulin G for immunosensors , 1991 .

[19]  A Ahluwalia,et al.  A comparative study of protein immobilization techniques for optical immunosensors. , 1992, Biosensors & bioelectronics.

[20]  R B Harris,et al.  Fiber-optic immunosensor for measurement of myoglobin. , 1997, Clinical chemistry.