Immunoassays using anti-allotypic monoclonal antibodies
AssigneeBayer Aktiengesellschaft
InventorEve H. Barlow
Life of this patent
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Abstract
The invention provides improved immunoassay techniques for detecting the presence of analytes in a liquid sample. The present immunoassay methods utilize anti-allotypic monoclonal antibodies as capture reagents for primary binding proteins specific for the analytes of interest. The monoclonal antibodies are highly specific for the allotypic determinants present on the primary binding protein. The use of anti-allotypic monoclonal antibodies as capture reagents provides improved levels of specificity and accuracy of the immunoassay, in part because interference from endogenous immunoglobulins in the sample is significantly reduced. The invention further provides anti-allotypic monoclonal antibodies.
Claims22
A method for determining the amount of analyte in a patient sample while reducing interference by endogenous immunoglobulins or complement proteins present in the sample, said method comprising:
- a. combining the patient sample with (i) a binding protein specific for said analyte, said binding protein having a specific allotype, and (ii) detectably labeled tracer molecules specific for said binding protein;
- b. incubating the combination formed in step (a) under conditions sufficient to permit the analyte and the labeled tracer molecules to bind to said binding protein;
- c. contacting the combination with a solid phase having immobilized thereon a capture antibody specific for the binding protein allotype wherein the capture antibody is obtained from hybridoma cell line ATCC 12349 or ATCC 12350;
- d. incubating the combination formed in step (c) under conditions sufficient to permit the binding protein to bind with the capture antibody;
- e. separating the solid phase from the sample;
- and
- f. detecting the amount of labeled tracer molecules bound to the solid phase;
- wherein said amount of labeled tracer detected is indicative of the amount of analyte present in the sample, and wherein the use of the allotype-specific antibody in step (c) reduces interference by endogenous immunoglobulins or complement proteins present in the sample.
A method for determining the amount of analyte in a patient sample, said method comprising:
- a. combining the patient sample with a primary monoclonal antibody and a tracer comprising acridinium-ester-labeled analyte molecules specific for said primary antibody;
- b. incubating the combination formed in step (a) under conditions sufficient to permit the analyte and the tracer to bind to said primary antibody;
- c. contacting the combination with a paramagnetic particle having immobilized thereon a capture anti-allotypic monoclonal antibody specific for the primary monoclonal antibody allotype wherein the capture antibody is obtained from hybridoma cell line ATCC 12349 or ATCC 12350;
- d. incubating the combination formed in step (c) under conditions sufficient to permit the primary monoclonal antibody to bind with the capture antibody;
- e. separating the solid phase from the sample, unbound tracer and unbound analyte;
- and
- f. detecting the amount of tracer bound to the solid phase;
- wherein said amount of labeled analyte molecules is indicative of the amount of analyte present in the sample.
Dependencies are inferred from the claim text.