C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/62 (2006.01) A01K 67/033 (2006.01) C07K 19/00 (2006.01) C12Q 1/02 (2006.01) G01N 33/52 (2006.01) G01N 33/567 (2006.01)
Patent
CA 2584108
Intramolecular biosensors are disclosed, including PBP-based biosensors, comprising a ligand binding domain fused to donor and fluorescent moieties that permit detection and measurement of Fluorescence Resonance Energy Transfer upon binding ligand. At least one of the donor and fluorescent moieties may be internally fused to the biosensor such that both ends of the internally fused fluorophore are fixed. In addition, methods of improving the sensitivity of terminally fused biosensors are provided. The biosensors of the invention are useful for the detection and quantification of ligands in vivo and in culture.
L'invention concerne des biocapteurs intramoléculaires, y compris des biocapteurs à protéines de liaison périplasmiques (PBP), qui comprennent un domaine de fixation de ligands fusionné à des fragments donneur et fluorescent permettant de détecter et de mesurer le transfert par résonance d'énergie de fluorescence après fixation du ligand. Au moins un des fragments donneur et fluorescent peut être fusionné au biocapteur de manière interne, de sorte que les deux extrémités du fluorophore fusionné de manière interne sont fixées. L'invention concerne en outre des procédés permettant d'améliorer la sensibilité de biocapteurs fusionnés à leurs extrémités. Les biocapteurs de l'invention sont utiles pour détecter et quantifier des ligands in vivo et en culture.
Fehr Marcus
Frommer Wolf B.
Looger Loren
Okumoto Sakiko
Carnegie Institution Of Washington
Gowan Intellectual Property
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