Heterogeneous assays for different analytes in a single biological sample
are performed simultaneously in a multiplexed assay that combines flow
cytometry with the use of magnetic particles as the solid phase and
yields an individual result for each analyte. The particles are
distinguishable from each other by characteristics that permit them to be
differentiated into groups, each group carrying an assay reagent bonded
to the particle surface that is distinct from the assay reagents of
particles in other groups. The magnetic particles facilitate separation
of the solid and liquid phases, permitting the assays to be performed by
automated equipment. Assays are also disclosed for the simultaneous
detection of antibodies of different classes and a common antigen
specificity or of a common class and different antigen specificities.
Each type is accomplished by immunological binding at the surfaces of two
distinct solid phases in a sequential manner with dissociation of the
binding and washing of the solid phase in between the binding steps.