Immobilized and stabilized loo_gfp fluorescent biosensors with an enhanced dynamic range
AssigneeThe Texas A&M University System
InventorsSarah Bondos, Rebecca Booth, Amanda Jons
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Abstract
Disclosed are leave-one-out GFP (LOO_GFP) biosensors immobilized in Ubx (Drosphila protein Ultrabithorax) that form immobilized and stabilized fluorescent protein biosensor systems (LOO_GFP-Ubx systems). These LOO_GFP-Ubx systems have an enhanced fluorescent signaling to noise ratio. Also disclosed are methods of making LOO_GFP-Ubx systems and methods of using such systems to detect analyte. In preferred embodiments, the LOO_GFP-Ubx system has the eighth β strand or eleventh f strand of OPT GFP left out.
Claims20
A composition comprising:
- a. a protein-fragment complementation assay wherein at least one component is immobilized with ultrabiothorax (Ubx) biomaterials;
- b, wherein the protein-fragment complementation assay comprises a fluorescent protein.
A method of manufacturing LOO_GFP-Ubx suitable for a biosensor comprising:
- a. co-expressing a LOO_GFP-Ubx fusion protein and a left-out peptide in a first host cell;
- b. extracting protein from the first host cell;
- c. incubating the extracted protein in a buffer reservoir to support film formation;
- d. pulling fibers from the film;
- e. air drying the fibers;
- and
- f. washing the fibers with a denaturing wash to remove the left-out peptide.
Dependencies are inferred from the claim text.