APPARATUS AND METHODS FOR MULTIANALYTE HOMOGENEOUS FLUOROIMMUNOASSAYS

摘要

Methods and apparatus for evanescent light fluoroimmunoassays are disclosed. The apparatus employs a planar waveguide (124) with an integral semi-cylindrical lens (502), and has multi-analyte features and calibration features, along with improved evanescent field intensity. A preferred embodiment of the biosensor (120) and assay method have patches (662) of capture molecules (720) each specific for a different analyte (210), disposed adjacent within a single reservoir (660). The capture molecules (720) are immobilized to the patches (662) on the waveguide surface by site-specific coupling of thiol groups on the capture molecules (720) to photo-affinity crosslinkers (706), which in turn are coupled to the waveguide surface (700) or to a non-specific-binding-resistant coating (702) on the surface (700). The patches (662, 664, 666) of different antibodies are produced by selectively irradiating a portion (714) of the waveguide surface (700) during the process of coupling the photo-affinity crosslinkers (706), the selective irradiation involving a mask (712), a laser light source, or the like.