Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays
First Claim
1. An optical substrate, said optical substrate comprising:
- an optical surface having a coated region coated with a coating providing a level of nonspecific protein binding to the optical surface which is below about 10% of the amount of specific protein binding, the coating including a coating material selected from the group consisting of polyethylene glycol (PEG), a hydrogel formed of polymethacryloyl polymers, a silyl-derivatized polyethyleneglycol, avidin-biotin-PEG, 3-aminopropyltriethoxy silane-Glu-PEG, and block copolymers, said block copolymers comprising a hydrophobic polymer block consisting essentially of hydrophobic residues adjacent at least one hydrophilic polymer block consisting essentially of hydrophilic residues.
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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.
192 Citations
26 Claims
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1. An optical substrate, said optical substrate comprising:
an optical surface having a coated region coated with a coating providing a level of nonspecific protein binding to the optical surface which is below about 10% of the amount of specific protein binding, the coating including a coating material selected from the group consisting of polyethylene glycol (PEG), a hydrogel formed of polymethacryloyl polymers, a silyl-derivatized polyethyleneglycol, avidin-biotin-PEG, 3-aminopropyltriethoxy silane-Glu-PEG, and block copolymers, said block copolymers comprising a hydrophobic polymer block consisting essentially of hydrophobic residues adjacent at least one hydrophilic polymer block consisting essentially of hydrophilic residues. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
Specification