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.