Homogeneous time resolved fluorescence based test system for Paramyxoviridae
AssigneeTibotec BVBA
InventorsDirk André E Roymans, Koenraad Lodewijk August van Acker, Inge Vereycken, Géry Karel Julia Dams
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Abstract
The present invention concerns a fluorescence resonance energy transfer based high throughput test system to measure the formation of the RSV F1 six-helix bundle. In a first embodiment the current invention relates to a homogeneous time resolved fluorescence-based test system comprising a first helical polypeptide consisting essentially of the sequence of IQN57 (SEQ ID NO: 1); a second helical polypeptide consisting essentially of the sequence of C45 (SEQ ID NO: 2) wherein said IQN57 is labeled with a light emitting fluorophore and said C45 is labeled with an ultra-violet excitable fluorophore.
Claims17
A homogeneous time resolved fluorescence-based test system comprising a first helical labeled polypeptide consisting of a sequence from the heptad-repeat 1 (HR1) region of Respiratory Syncytial Virus (RSV);
- and a second helical labeled polypeptide consisting of a sequence from the heptad-repeat 2 (HR2) region of RSV, wherein the first helical polypeptide consists of the sequence of IQN57 (SEQ ID NO:
- 1), and the second helical polypeptide consists of the sequence of C45 (SEQ ID NO:
- 2).
A method for identifying a compound that interferes with the formation of the RSV six-helix bundle of F1 comprising:
- providing a first helical polypeptide consisting of the sequence of IQN57 (SEQ ID NO:
- 1);
- providing a second helical polypeptide consisting of the sequence of C45 (SEQ ID NO:
- 2) wherein said IQN57 is labeled with a light emitting fluorophore and said C45 is labeled with an ultra-violet excitable fluorophore;
- providing a test composition comprising the compound;
- measuring the degree of complex formation between the first helical polypeptide and the second helical polypeptide in the presence of the test composition comprising the compound using fluorescence resonance energy transfer;
- and
- comparing the measured degree of complex formation to the degree of complex formation between the first helical polypeptide and the second helical polypeptide in the absence of the test composition comprising the compound to identify the compound that interferes with the formation of the RSV six-helix bundle of F1.
A method for identifying the mechanism of inhibition of the formation of the RSV six-helix bundle of F1 comprising:
- providing a first helical polypeptide consisting of the sequence of IQN57 (SEQ ID NO:
- 1);
- providing a second helical polypeptide consisting of the sequence of C45 (SEQ ID NO:
- 2) wherein said IQN57 is labeled with a light emitting fluorophore and said C45 is labeled with an ultra-violet excitable fluorophore;
- providing a test composition comprising a compound that interferes with the formation of the six-helix bundle of F1;
- measuring the degree of complex formation between the first helical polypeptide and the second helical polypeptide in the presence of the test composition comprising a compound that interferes with the formation of the six-helix bundle of F1 using fluorescence resonance energy transfer;
- and
- comparing the measured degree of complex formation to the degree of complex formation between the first helical polypeptide and the second helical polypeptide in the absence of the test composition comprising the compound that interferes with the formation of the six-helix bundle of F1 to identify the mechanism of inhibition of the formation of the RSV six-helix bundle of F1.
Dependencies are inferred from the claim text.