Real time detection of intermolecular interaction
AssigneeTrellis Bioscience, LLC
InventorLawrence M. Kauvar
Life of this patent
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Abstract
Intermolecular binding can be detected by formation of a “paratope” which results in an immediate generation of a signal. The substances to be tested for interaction are bound to demitopes, wherein said demitopes are components of a paratope which binds a reporter which provides said signal when bound. Known interactions measured in this way can also be employed to screen for compounds which interfere with the interactions. In addition to testing for individual interactions, the interaction of a compound with a library or library×library interactions can also be determined and the effect of potentially interfering substances evaluated.
Claims29
A method to determine whether a first substance interacts with a second substance which method comprises
- contacting said first substance coupled to a first demitope with said second substance coupled to a second demitope, in the presence of a reporter wherein said first and second demitopes, when assembled, form a paratope which binds said reporter, and wherein said reporter generates an immediate detectable signal when bound to the paratope, and
- determining the presence, absence or magnitude of the signal produced;
- wherein the presence of the signal indicates said first and second substances interact and the absence of signal indicates the first and second substance do not interact, or
- the magnitude of the signal is a measure of the affinity of interaction of said first substance and said second substance.
A method to identify a compound which interferes with the interaction of a first substance with a second substance which method comprises
- contacting, in the absence of a candidate compound, said first substance coupled to a first demitope with said second substance coupled to a second demitope in the presence of a reporter, wherein said first and second demitopes when assembled, form a paratope which binds said reporter and wherein said reporter generates an immediate detectable signal when bound to the paratope, and wherein said first substance and said second substance interact to generate an immediate detectable signal;
- contacting, in the presence of a candidate compound, said first substance coupled to a first demitope with said second substance coupled to a second demitope in the presence of a reporter, wherein said first and second demitopes when assembled, form a paratope which binds said reporter and wherein said reporter generates an immediate detectable signal when bound to the paratope, and wherein said first substance and said second substance interact to generate an immediate detectable signal;
- determining the presence, absence or magnitude of the signal produced in each case;
- comparing the signal produced in the presence of said candidate compound with the signal produced in the absence of the candidate compound;
- wherein a reduction in the magnitude of the signal produced or the abolition of the signal produced in the presence of said compound as compared to the signal in the absence of said compound identifies said compound as interfering with the interaction of said first and second substance.
A compound of the formula X—A, wherein X is a substance to be tested for interaction and A is a demitope selected from the group consisting of a variable light chain, a variable heavy chain, and a portion of a receptor.
A kit for the determination of interaction of substances, which kit comprises, in separate containers, a first demitope, a second demitope complementary to the first demitope and, optionally, a reporter reagent which binds to the paratope formed by the interaction of the first and second demitope to provide an immediate signal.
Dependencies are inferred from the claim text.