Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays
First Claim
1. Apparatus for homogeneous solid-state fluorescence immunoassays, comprising a biosensor havinga planar waveguide having first and second parallel plane surfaces with a surrounding edge extending between said first and second surfaces, said edge having a receiving region for receiving light, and at least one of said surfaces having a plurality of capture molecules immobilized thereon, said capture molecules each having a binding site which selectively binds an analyte, anda semi-cylindrical lens located adjacent to the edge of said planar waveguide.
2 Assignments
0 Petitions
Accused Products
Abstract
Methods and apparatus for evanescent light fluoroimmunoassays are disclosed. The apparatus employs a planar waveguide with an integral semi-cylindrical lens, and has multi-analyte features and calibration features, along with improved evanescent field intensity. A preferred embodiment of the biosensor and assay method have patches of capture molecules each specific for a different analyte disposed adjacent within a single reservoir. The capture molecules are immobilized to the patches on the waveguide surface by site-specific coupling of thiol groups on the capture molecules to photo-affinity crosslinkers which in turn are coupled to the waveguide surface or to a non-specific-binding-resistant coating on the surface. The patches of different antibodies are produced by selectively irradiating a portion of the waveguide surface during the process of coupling the photo-affinity crosslinkers the selective irradiation involving a mask, a laser light source, or the like.
-
Citations
26 Claims
-
1. Apparatus for homogeneous solid-state fluorescence immunoassays, comprising a biosensor having
a planar waveguide having first and second parallel plane surfaces with a surrounding edge extending between said first and second surfaces, said edge having a receiving region for receiving light, and at least one of said surfaces having a plurality of capture molecules immobilized thereon, said capture molecules each having a binding site which selectively binds an analyte, and a semi-cylindrical lens located adjacent to the edge of said planar waveguide.
-
17. A method of performing a solid-state fluoroimmunoassay in which a capture molecule specific for an analyte is coupled to an optical substrate, and in which binding of the analyte to the capture molecule results in a fluorescent signal corresponding to the presence of the analyte in a sample, comprising the steps of:
-
providing a biosensor comprising a planar waveguide having a surrounding edge extending between first and second parallel surfaces, a semi-cylindrical lens located adjacent to the edge of said planar waveguide, wherein at least one of said surfaces has a plurality of capture molecules immobilized thereon, said capture molecules each having a binding site which selectively binds an analyte; providing a light source disposed to deliver a light beam into the waveguide through the semi-cylindrical lens; providing detection means disposed for detecting fluorescent light which impinges on a plane parallel to and displaced from said waveguide surfaces; contacting the waveguide surface having the capture molecules immobilized thereon with a solution comprising a buffer, a plurality of tracer molecules capable of binding to the capture molecule in the presence of the analyte and of emitting a fluorescent signal upon stimulation with the light beam, and an unknown amount of analyte molecules; incubating the waveguide surface with the contacting solution to permit binding of the tracer molecules to the analyte molecules and the capture molecules; operating the light source to produce evanescent light from the waveguide to stimulate bound tracer molecules; operating the detection means to detect the fluorescent signals emitted by the bound tracer molecules; and analyzing the fluorescent signals emitted to determine the amount of the analyte in the test sample. - View Dependent Claims (18, 19, 20)
-
-
21. Apparatus for homogeneous solid-state fluorescence immunoassays, comprising:
-
a planar waveguide having first and second parallel plane surfaces with a surrounding edge extending between said first and second surfaces, said edge having a receiving region for receiving light, and at least one of said surfaces having a plurality of capture molecules immobilized thereon, said capture molecules each having a binding site which selectively binds an analyte; a semi-cylindrical lens located adjacent to the edge of said planar waveguide a light source providing a light beam in a desired wavelength range and positioned to aim said beam into said receiving region of said waveguide; one or more reservoirs formed on at least one of said waveguide surfaces, each said reservoir configured for containing a sample solution comprising a buffer, a plurality of molecules of said analyte, and a plurality of tracer molecules capable of emitting fluorescent light upon stimulation with evanescent light resulting from said beam; light collection means oriented to non-evanescently collect a portion of said fluorescent light which impinges on a plane parallel to and displaced from the waveguide surface; and detection means optically associated with said light collection means for detecting and quantitating said fluorescent light to produce a fluorescence signal reflective of said stimulation of said tracer molecules. - View Dependent Claims (22, 23, 24, 25, 26)
-
Specification