Quantitative multiplex detection of pathogen biomarkers
View Patent ↗The present invention addresses the simultaneous detection and quantitative measurement of multiple biomolecules, e.g., pathogen biomarkers through either a sandwich assay approach or a lipid insertion approach. The invention can further employ a multichannel, structure with multi-sensor elements per channel.
1. A method of simultaneously detecting and quantitatively measuring multiple biomolecules characterized by having a lipophilic portion of sufficient size and chemical composition for insertion into a lipid bilayer, the method comprising:
contacting a target sample containing multiple differing target biomolecules with an assay element including a substrate, a patterned waveguide upon said substrate, a lipid bilayer upon the patterned waveguide, a reference biomolecule including an attached inorganic fluorescent reporter molecule, the reference biomolecule attached to said lipid bilayer as an assay element standard;
maintaining the target sample together with the assay element for a time sufficient for a insertion event to occur between target biomolecules characterized by having a lipophilic portion of sufficient size and chemical composition for insertion into a lipid bilayer within the sample and the lipid bilayer;
washing excess biological target sample from contact with the assay element;
contacting the assay element with an amount of at least two additional recognition ligands each attached to a different inorganic fluorescent reporter molecule, each additional recognition ligand binds to a specifically targeted biomolecule;
washing excess of the at least two additional recognition ligands from contact with the assay element;
interrogating the assay element with excitation light from the waveguide; and,
obtaining an output correlated to detection and quantitative measurement of target biomolecules within the target sample.
2. An assay element comprising:
a substrate;
a patterned waveguide upon said substrate;
a lipid bilayer upon the patterned waveguide; and
a reference biomolecule including an attached inorganic fluorescent reporter molecule, the reference biomolecule attached to said lipid bilayer as an assay element standard.
3. A kit for simultaneous detection and quantitative measurement of multiple biomolecules characterized by having a lipophilic portion of sufficient size and chemical composition for insertion into a lipid bilayer, the kit comprising: (1) a assay element including a substrate, a patterned waveguide upon said substrate, a lipid bilayer upon the patterned waveguide, a reference biomolecule including an attached inorganic fluorescent reporter molecule, the reference biomolecule attached to said lipid bilayer as an assay element standard; and (2) an amount of at least two additional recognition ligands each attached to a different reporter molecule and each additional recognition ligand binds to one specifically targeted biomolecule.