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AssigneeBristol-Myers Squibb Company
InventorsSusan B. Roberts, Roger Hochoon Pak
Estimating the term…
The present invention provides an isolated, functionally-active protein that catalyzes cleavage of a gamma-secretase substrate. The functional activity of the isolated protein suggests that the isolated protein includes gamma-secretase. In one embodiment, the isolated gamma-secretase protein is associated with PS1. The present invention also relates to homogeneous methods for monitoring cleavage of β-amyloid precursor protein (βAPP) by gamma-secretase, wherein the steps of of isolating and retrieving cleavage products have been eliminated. Cleavage can be detected by binding a pair of fluorescent adducts to the gamma-cleaved βAPP fragment. Preferably, a first fluorescent adduct binds to the carboxy-terminal end of the gamma-cleaved βAPP fragment, with substantially no cross-reactivity to uncleaved βAPP or to other types of gamma-cleaved βAPP fragments, while a second fluorescent adduct binds to a portion within the amino-terminal region on the gamma-cleaved βAPP fragment. Detection of binding to the gamma-cleaved βAPP fragment is determined by monitoring the fluorescent energy transfer between the adducts.
An in vitro homogenous method of detecting cleavage of β-amyloid precursor protein (βAPP) by gamma-secretase, said method comprising detecting binding of a gamma-cleaved βAPP fragment with a pair of fluorescent adducts, wherein a first fluorescent adduct binds specifically to the carboxy-terminal end of the gamma-cleaved βAPP fragment with substantially no cross-reactivity to uncleaved βAPP or to other types of gamma-cleaved βAPP fragments, and wherein a second fluorescent adduct binds to the gamma-cleaved βAPP fragment within an amino acid sequence which corresponds to amino acids 1-31 of β-amyloid peptide (Aβ), and wherein excitation of one of the fluorescent adducts provides a detectable transfer of fluorescent energy to the other fluorescent adduct.
An in vitro homogenous method for determining the presence of β-amyloid peptide (Aβ), in a sample said method comprising:
An in vitro homogenous method for determining the presence of β-amyloid peptide Aβ-40 in a sample, said method comprising:
An in vitro homogenous method of detecting cleavage of β-amyloid precursor protein (βAPP) by gamma-secretase, said method comprising detecting binding of a 6 kDa C-terminal fragment with a pair of fluorescent adducts, wherein a first fluorescent adduct binds to the amino-terminal end of the 6 kDa C-terminal fragment with substantially no cross-reactivity to uncleaved βAPP or to other types of gamma-cleaved βAPP fragments, and wherein a second fluorescent adduct binds to a portion within the carboxy-terminal region of the 6 kDa C-terminal fragments and wherein excitation of one of the fluorescent adducts provides a detectable transfer of energy to the other fluorescent adduct.
An in vitro homogenous method of detecting cleavage of β-amyloid precursor protein (βAPP) by gamma-secretase, comprising the steps of:
An in vitro homogenous method of screening for inhibitors of gamma-secretase in β-amyloid precursor protein (βAPP), said method comprising the steps of:
An in vitro homogenous method of screening for inhibitors of gamma-secretase in β-amyloid precursor protein (βAPP), said method comprising the steps of:
Dependencies are inferred from the claim text.