Fluorescent fusion polypeptides and methods of use

摘要

Embodiments of the present invention provide for the facile generation of a stable recombinant fusion polypeptides with intrinsic fluorescent properties. The recombinant antibodies may be suitable for qualitative and/or quantitative immunofluorescence analysis. Generally, the fluorescent polypeptides include a fluorescent domain comprising a C-terminus and an N-terminus; a first antibody domain covalently linked to the C-terminus; and a second antibody domain covalently linked to the N-terminus.

主权项

1. A method of detecting an analyte comprising:
providing a sample that comprises the analyte; contacting the sample with a polypeptide that comprises:
a fluorescent domain comprising:
a monomeric fluorescent polypeptide comprisinga C-terminus; andan N-terminus;a first antibody domain covalently linked to the C-terminus of the fluorescent domain, wherein the first antibody domain comprises a variable light chain (VL) or a variable heavy chain (VH); anda second antibody domain covalently linked to the N-terminus of the fluorescent domain, wherein the second antibody domain comprises a variable light chain (VL) or a variable heavy chain (VH);wherein the N-terminus of first antibody domain and the C-terminus of the second antibody domain are separated by a distance of no less than 30 Å and no more than 40 Å, wherein at least one of the first antibody domain and the second antibody domain specifically binds to the analyte; removing unbound polypeptides; and detecting a fluorescent signal produced by the polypeptide specifically bound to the analyte, thereby detecting presence of the analyte in the sample.