Label-free monitoring of antibody–antigen interactions using optical microchip biosensors

Abstract

A rapid, label-free optical biosensor system for sensitive monitoring of bio-molecular interactions in real-time is presented. SpectroSens™ sensor chips are based on integrated planar Bragg gratings sensitive to localised changes in refractive index. Bio-molecule recognition is imparted by functionalisation of the sensing surface with antibodies against targets of interest. In this study, antibodies against selected proteins were oriented with recombinant Protein A/G, which was covalently immobilised to the sensor chip via amine coupling to a glutaraldehyde-activated silane layer. Immunoassays for the detection of rabbit IgG and ovalbumin proteins as model antibody–antigen interaction systems were performed. Binding of complementary antigens to respective antibody-functionalised sensors manifested as changes in wavelength of light reflected from the optical sensors. Quantitative binding kinetics with detection sensitivities in the mid ng/ml range were obtained for both antigens using this planar, two-dimensional surface coating. Data presented demonstrate the suitability of SpectroSens™ sensors as a valuable tool in life science research and development for monitoring bio-specific interactions, protein concentration determination and antibody selection; the optical integration and analytical characteristics of these sensors suggest that they may find numerous applications in bio-pharmaceutical development and clinical diagnostics.