Antibodies to complexes of ligand receptors and ligands and their utility in ligand-receptor assays
AssigneeBiosite Diagnostics, Inc.
InventorsKenneth F. Buechler, Gunars E. Valkirs
Life of this patent
Estimating the term…
Abstract
Methods and test devices for detecting the presence or amount of target ligand in non-competitive sandwich ligand-receptor assay processes. Antibodies which bind to the complex of ligand receptor and target ligand but do not bind significantly to the ligand receptor and which bind the target ligand with substantially less affinity than the complex are taught and their uses described. These assays can be used to eliminate the "hook" effect in non-competitive sandwich assays. Furthermore, the antibodies are selected and assay methods described so that, as a result of the assay process, no detectable response is observed due to the binding of antibody and ligand receptor in the absence of target ligand.
Claims40
A device for performing an assay to detect the presence of a target ligand in a sample, said device comprising:
- a first receptor immobilized to a surface of the device, wherein the affinity of the first receptor for a complex of a target ligand and a second receptor is at least ten times as great as the affinity of the first receptor for the target ligand alone, and wherein said device is adapted and arranged such that the assay, when performed, is essentially unaffected by a hook effect.
A device for performing an assay to detect the presence of a target ligand in a sample, said device comprising:
- a first receptor immobilized to a surface of the device, said first receptor able to bind the target ligand to form a complex;
- a second receptor, said second receptor capable of binding the complex, wherein the affinity of the second receptor for the complex is at least ten times as great as the affinity of the second receptor for the target ligand;
- a means for contacting the second receptor with the complex when complex has been formed, wherein said device is adapted and arranged such that the assay, when performed, is essentially unaffected by a hook effect.
A device for performing an assay to detect the presence of a target ligand in a sample, said device comprising:
- a first receptor coupled to a signal development element, said first receptor capable of binding the target ligand to form a complex;
- a second receptor, said second receptor capable of binding the complex, wherein the affinity of the second receptor for the complex is at least ten times as great as the affinity of the second receptor for the target ligand;
- and,
- a third receptor immobilized to a surface of the device, said third receptor capable of binding the second receptor when it is bound to the complex, wherein said device is adapted and arranged such that the assay, when performed, is essentially unaffected by a hook effect.
A kit for use in conducting an assay that is essentially unaffected by a hook effect, where the assay determines the presence or amount of a target ligand, said kit comprising:
- a first receptor, said first receptor capable of binding the target ligand to form a complex;
- and,
- a second receptor, said second receptor capable of binding the complex, wherein the affinity of the second receptor for the complex is at least ten times as great as the affinity of the second receptor for the target ligand;
- wherein said kit is adapted and arranged such that the assay, when performed, is essentially unaffected by a hook effect.
A test device for performing an assay to determine the presence or amount of at least one target ligand in a fluid sample suspected of containing said target ligand, said target ligand capable of binding to a ligand receptor conjugate and to an antibody immobilized to a surface of the device, said device further comprising:
- a. a means for contacting said fluid sample with said ligand receptor conjugate and the immobilized antibody, said antibody capable of binding the complex of said target ligand and said ligand receptor conjugate, the binding affinity of said antibody for said complex being at least a factor of 10 greater than the affinity of said antibody for said target ligand, wherein no detectable assay response results from the binding of antibody and ligand receptor conjugate in the absence of target ligand, wherein the amount of ligand receptor conjugate bound to antibody is related to the amount of target ligand in the fluid sample;
- b. a means for detecting said complex bound to said antibody;
- and,
- c. a means for relating the presence or amount of complex detected to the presence or amount of said target ligand in said fluid sample, wherein said device is adapted and arranged such that the assay, when performed, is essentially unaffected by a hook effect.
A test device for performing an assay to determine the presence or amount of at least one target ligand in a fluid sample suspected of containing said target ligand, said target ligand capable of binding to a ligand receptor immobilized on a surface of the device and an antibody conjugate, said device further comprising:
- a. a means for contacting said fluid sample with said immobilized ligand receptor and said antibody conjugate, said antibody conjugate capable of binding the complex of said target ligand and said ligand receptor, the binding affinity of said antibody conjugate for said complex being at least a factor of 10 greater than the affinity of said antibody conjugate for said target ligand, wherein no detectable assay response results from the binding of antibody conjugate and ligand receptor in the absence of target ligand, and wherein the amount of ligand receptor bound to antibody conjugate is related to the amount of target ligand in the fluid sample;
- b. a means for detecting said complex bound to said antibody conjugate;
- and,
- c. a means for relating the presence or amount of complex detected to the presence or amount of said target ligand in said fluid sample, wherein said device is adapted and arranged such that the assay, when performed, is essentially unaffected by a hook effect.
A test device for performing an assay to determine the presence or amount of at least one target ligand in a fluid sample suspected of containing said target, said target ligand capable of binding to a ligand receptor and to an antibody conjugate, said device comprising:
- a. a means for contacting said fluid sample with said ligand receptor and said antibody conjugate, said antibody conjugate capable of binding a complex of said target ligand and said ligand receptor, the binding affinity of said antibody conjugate for said complex being at least a factor of 10 greater than the affinity of said antibody conjugate for said target ligand, wherein no detectable assay response results from the binding of antibody conjugate and ligand receptor in the absence of target ligand, wherein the amount of antibody conjugate bound to ligand receptor is related to the amount of target ligand in the fluid sample;
- b. a receptor immobilized to a surface of the device, the immobilized receptor capable of binding ligand receptor when ligand receptor is bound to the target ligand having formed a complex;
- c. a means for detecting said antibody conjugate bound to said complex;
- and,
- d. a means for relating the presence or amount of complex detected to the presence or amount of target ligand in said fluid sample, wherein said device is adapted and arranged such that the assay, when performed, is essentially unaffected by a hook effect.
Dependencies are inferred from the claim text.