A label-free porous alumina interferometric immunosensor.

Anodization of Al is used to produce optically smooth porous alumina (Al(2)O(3)) films with pores approximately 60 nm in diameter and approximately 6 mum deep. The capture protein, protein A, is adsorbed to the pore walls by noncovalent, electrostatic interactions, and thin film interference spectroscopy is used to detect binding of immunoglobulin (IgG). The porous alumina films are stable against corrosion and dissolution in aqueous media at pH 7, allowing quantitative monitoring of steady-state and time-resolved biomolecular binding. The bare porous Al(2)O(3) surface displays a significantly greater affinity for protein A than for IgG. The known species specificity of protein A binding to IgG is confirmed; the protein-A-modified sensor responds to IgG derived from rabbit, but not chicken (IgG/IgY). A "cascaded", or multiprobe sensing approach, is demonstrated, in which a specific target, sheep IgG, is administered to a sample modified with a protein A/rabbit anti-sheep IgG assembly. Binding measurements are confirmed by fluorescence microscopy using fluorescein-labeled IgG.