Quantitative multiplex detection of pathogen biomarkers
AssigneeLos Alamos National Security, LLC
InventorsHarshini Mukundan, Hongzhi Xie, Basil I. Swanson, Jennifer Martinez, Wynne K. Grace
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Abstract
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.
Claims3
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 adapted for binding 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.
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.
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 adapted for binding to one specifically targeted biomolecule.
Dependencies are inferred from the claim text.