IIBYT   23944
INSTITUTO DE INVESTIGACIONES BIOLOGICAS Y TECNOLOGICAS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Binding kinetics between soluble β-Gal and anti-β-Gal immobilized on chemically nanopatterned surfaces with fractal topography
Autor/es:
PEDRO D. CLOP; GERARDO R. MARCHESINI; MARIA A. PERILLO
Lugar:
Carlos Paz
Reunión:
Congreso; XLII Reunion Anual de la Sociedad Argentina de Biofì...; 2013
Institución organizadora:
Sociedad Argentina de Biofîsica
Resumen:
The present work was aimed
at providing experimental support to the correlation between topographic and
kinetic dimensions of protein function we described previously. Through
electronic and colloidal nanolitographic techniques we produced surfaces
exhibiting a fractal-like pattern with a pre-determined topographic dimension of
domains capable to bind proteins in a covalent manner (AbAntiβ -Gal).
These surfaces were used as sensors to reversibly bind β-Gal and enabled the
kinetic study of AbAnti β-Gal -β-Gal complex formation by surface
plasmon resonance (SPR) spectroscopy. Compared with the behavior of control
sensors (homogeneous topography), the Ag-Ab binding kinetics in sensors produced
by nanolithography showed higher capacity and broader dispersion of binding
sites characterized by a more diffuse attractor in the kd vs. Kd
phase space. Our results contributed to the development
of a nano-biosensor for β-Gal, to a strategy to enhance the sensitivity of a SPR
sensor and to endorse the cross-talk between supramolecular structure and
dynamics in heterogenous systems. Additionally, a catalytic activity could be
measured for the first time by a SPR based method where the insoluble product
of a β-Gal catalyzed reaction interfered with the evanescent field on the
sensor´s surface.