Patent ID: 7674632
Filing Date: 2010-03-09
Classification: G01N,Y10T

Abstract:
1. A method for multiplexed homogeneous clinical immunoassay for detecting antibodies to specific antigens using microparticles for diagnostic purposes comprising: A. providing a multiplexed microparticle suspension having a plurality of individually identifiable subsets of microparticles having a diameter of at least approximately 1.0 microns and including a plurality of test subsets of microparticles each having a specific antigen positioned thereon which is capable of binding with a specific target antibody and including at least two calibration subsets of microparticles having a first binding partner thereon at known concentrations on each of said calibration subsets which is capable of binding with human antibodies and further including at least one verification subset of microparticles having a second binding partner thereon at a known concentration level; B. first mixing of the multiplexed suspension of microparticles with a human serum sample to allow binding of specific target antibodies to the specific antigens on each of said test subsets of microparticles and to facilitate binding of human antibodies to said first binding partners on said calibration subsets of microparticles; C. first incubating of the mixture of the human serum sample and the multiplexed microparticle suspension together; D. providing of a tagging component including an attaching component capable of binding with respect to human antibodies and with respect to the second binding partner and also including an indicating component conjugated with the attaching component which is capable of being detected; E. second mixing of the tagging component with the first incubated mixture of human serum sample and multiplexed microparticle suspension to facilitate binding of the tagging component with respect to antibodies which are bound to microparticles of the test subsets and calibration subsets and with respect to the second binding partner on the verification subset; F. second incubating of the tagging component with the first incubated mixture of human serum sample and the multiplexed microparticle suspension by maintaining thereof at a predefined temperature for a predefined period of time; G. measuring of the tagging component bound to microparticles of each test subset, calibration subset and verification subset of microparticles within the second incubated mixture; H. defining a mathematical algorithm expressing the mathematical relationship between the measured tagging component and the concentration level of antibody bound to each of the microparticles by measuring the tagging component found attached to microparticles of each of the calibration subsets thereof; I. verifying the defined mathematical algorithm by measuring the tagging component found on the verification subset of microparticles; J. measuring the amount of tagging component bound to each test subset of microparticles; and K. determining the concentration level of specific target antibodies within the human serum sample by applying the verified mathematical algorithm to the measured amount of tagging component bound to each test subset of microparticles.