Multiplex flow assays preferably with magnetic particles as solid phase

A method for the analysis of a sample to simultaneously yet individually detect antibodies of different classes that have a single common antigen specificity, said method comprising:

• contacting said sample with a first solid phase coated with said antigen to cause antibodies in said sample to bind to said antigen;
• washing said first solid phase to isolate said first group from unbound species;
• suspending said first solid phase thus washed in a liquid suspending medium and releasing said antibodies from said antigen into said medium to form a supernatant containing unbound multiple-class antibodies specific to said antigen;
• isolating said supernatant from said first solid phase and contacting said supernatant with a second solid phase, said second solid phase comprising one or more portions each coated with an immunological binding member with specific binding affinity for a single antibody class, and each portion being capable of differentiation from other such portions;
• and
• individually detecting the occurrence of immunological binding between said antibody classes and said immunological binding members while differentiating between said portions.

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 supernatant containing antibodies of a said selected class but of multiple antigen specificities;
• isolating said supernatant from said first solid phase and contacting said supernatant with a second solid phase, said second solid phase comprising one or more portions each coated with an immunological binding member with specific binding affinity for antibodies of a single antigen specificity that is distinct from antigen specifities of other portions, and each portion being capable of differentiation from other such portions;
• and
• individually detecting the occurrence of immunological binding between said antibodies and said immunological binding members while differentiating between said portions.