Patent ID: 6960478
Filing Date: 2005-11-01
Classification: G01N,Y10S,Y10T

Abstract:
1. A method for the analysis of a sample to simultaneously yet individually detect antibodies of different antigen specificities that are of a single common immunoglobulin class, said method comprising: contacting said sample with a first solid phase coated with class-specific antibody having specific binding affinity for a single selected class of antibodies to cause antibodies in said sample of said single selected class to be immobilized to said first solid phase through immunological binding to said class-specific antibody coated thereon, washing said first solid phase to isolate said first solid phase from unbound species; contacting said first solid phase thus washed with a liquid medium and releasing said antibodies of said single selected class from said first solid phase by dissociating said immunological binding to form a supematant containing antibodies of a said selected class but of multiple antigen specificities; isolating said supematant from said first solid phase and contacting said supematant with a second solid phase, said second solid phase comprising one or more portions each having coupled thereto an immunological binding member with specific binding affinity for antibodies of a single antigen specificity that is distinct from antigen specificities of other portions, and each portion being capable of differentiation from other such portions wherein the second solid phase comprises microparticles of magnetically responsive material, the sizes of said microparticles varying in size over a range that is an aggregate of a plurality of subranges, each subrange distinguishable from other subranges of said aggregate by flow cytometry and by the immunological binding member coupled thereto, said microparticles being suitable for use in a multiplex assay procedure that includes the use of flow cytometry; and individually detecting the occurrence of immunological binding between said antibodies and said immunological binding members while differentiating between said portions.