Back to results
US 5,480,792 AGranted
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.
Claims21
Method for determining the presence of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with a ligand receptor conjugate and an antibody wherein the binding affinity of said antibody for the complex of said target ligand and said ligand receptor conjugate is at least a factor of 10 greater than the affinity of said antibody for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor conjugate, and wherein no detectable assay response results from the binding of antibody and ligand receptor conjugate in the absence of target ligand and the amount of ligand receptor conjugate bound to antibody is related to the amount of target ligand in the fluid sample;
- b. measuring the amount of said complex bound to said antibody;
- c. relating said amount to the presence of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to a hook-effect at high ligand concentration.
1 more element
Method for determining the presence of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with a ligand receptor conjugate and an antibody wherein the binding affinity of said antibody for the complex of said target ligand and said ligand receptor conjugate is at least a factor of 10 greater than the affinity of said antibody for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor conjugate and wherein no detectable assay response results from the binding of antibody and ligand receptor conjugate in the absence of target ligand and the amount of ligand receptor conjugate bound to antibody is related to the amount of target ligand in the fluid sample;
- b. contacting the fluid from step (a) with means for removing said antibody and moieties bound to it from said fluid;
- c. measuring the amount of said complex bound to said antibody;
- d. relating said amount to the presence of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to a hook-effect at high ligand concentration.
2 more elements
Method for determining the presence of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with a ligand receptor and an antibody conjugate wherein the binding affinity of said antibody conjugate for the complex of said target ligand and said ligand receptor is at least a factor of 10 greater than the affinity of said antibody conjugate for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor and wherein no detectable assay response results from the binding of antibody conjugate and ligand receptor in the absence of target ligand and the amount of antibody conjugate bound to ligand receptor is related to the amount of target ligand in the fluid sample;
- b. measuring the amount of said antibody conjugate bound to said complex;
- c. relating said amount to the presence of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to a hook-effect at high ligand concentration.
1 more element
Method for determining the presence of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with a ligand receptor and an antibody conjugate wherein the binding affinity of said antibody conjugate for the complex of said target ligand and said ligand receptor is at least a factor of 10 greater than the affinity of said antibody conjugate for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor, and wherein no detectable assay response results from the binding of antibody conjugate and ligand receptor in the absence of target ligand and the amount of ligand receptor bound to antibody is related to the amount of target ligand in the fluid sample;
- b. contacting the fluid from step (a) with a means removing ligand receptor and moieties bound to it from said fluid;
- c. measuring the amount of said antibody conjugate bound to said complex;
- d. relating said amount to the presence of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to a hook-effect at high ligand concentration.
2 more elements
Method for determining the amount of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with a ligand receptor conjugate and an antibody wherein the binding affinity of said antibody for the complex of said target ligand and said ligand receptor conjugate is at least a factor of 10 greater than the affinity of said antibody for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor conjugate and wherein no detectable assay response results from the binding of antibody and ligand receptor conjugate in the absence of target ligand and the amount of ligand receptor conjugate bound to antibody is related to the amount of target ligand in the fluid sample;
- b. measuring the amount of said complex bound to said antibody;
- c. relating said amount to the amount of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to a hook-effect at high ligand concentration.
1 more element
Method for determining the amount of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps
- a. contacting said fluid sample with a ligand receptor conjugate and an antibody wherein the binding affinity of said antibody for the complex of said target ligand and said ligand receptor conjugate is at least a factor of 10 greater than the affinity of said antibody for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor conjugate and wherein no detectable assay response results from the binding of antibody and ligand receptor conjugate in the absence of target ligand and the amount of ligand receptor conjugate bound to antibody is related to the amount of target ligand in the fluid sample;
- b. contacting the fluid from step (a) with means for removing said antibody and moieties bound to it from said fluid;
- c. measuring the amount of said complex bound to said antibody;
- d. relating said amount to the amount, of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to a hook-effect at high ligand concentration.
2 more elements
Method for determining the amount of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with a ligand receptor and an antibody conjugate wherein the binding affinity of said antibody conjugate for the complex of said target ligand and said ligand receptor is at least a factor of 10 greater than the affinity of said antibody conjugate for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor, and wherein no detectable assay response results from the binding of antibody conjugate and ligand receptor in the absence of target ligand and the amount of antibody conjugate bound to ligand receptor is related to the amount of target ligand in the fluid sample;
- b. measuring the amount of antibody conjugate bound to said complex;
- c. relating said amount to the amount of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to a hook-effect at high ligand concentration.
1 more element
Method for determining the amount of at least one target ligand in a fluid sample suspected of containing said target ligand comprising the steps of:
- a. contacting said fluid sample with a ligand receptor and an antibody conjugate wherein binding affinity of said antibody conjugate for the complex of said target ligand and said ligand receptor is at least a factor of 10 greater than the affinity of said antibody conjugate for said target ligand when measured in the presence of a concentration of said target ligand that is substantially greater than the concentration of said ligand receptor, and wherein no detectable assay response results from the binding of antibody conjugate and ligand receptor in the absence of target ligand and the amount of antibody conjugate bound to ligand receptor is related to the amount of target ligand in the fluid sample;
- b. contacting the fluid from step (a) with a means for removing ligand receptor and moieties bound to it from said fluid;
- c. measuring the amount of antibody conjugate bound to said complex;
- d. relating said amount to the amount of said target ligand in said fluid sample;
- wherein sandwich immunoassays of said ligand are subject to hook-effect at high ligand concentration.
2 more elements
Dependencies are inferred from the claim text.