Flow based assays for therapeutics

A flow cytometry method of analyzing an immune reaction to a therapeutic antibody of a patient in vitro, the therapeutic antibody being capable of multiple modes of action so that it will bind a cell depletion surface marker to effect lysis and enable engagement of effector cells, the method comprising:

• exposing a whole blood sample from a patient to the therapeutic antibody in dry reagent form;
• combining exposed blood cells from the whole blood sample with a panel of a plurality of labeled reporters comprising at least two labeled reporters selected from the group consisting of a labeled reporter directed against a target cell depletion surface marker, a labeled reporter directed against an activation surface marker expressed on white blood cell populations, and a labeled reporter directed against cytokine production by white blood cells to thereby simultaneously produce assay signals generated by the at least two labeled reporters;
• subjecting the blood cell sample mixture to flow cytometry and measuring signals generated by each of the plurality of labeled target cells and labeled cytokine;
• and
• combining the measured signals into an assay output for flow cytometric multiparameter analysis, the assay output indicative of the patient's immune reaction to the therapeutic antibody and the assay output providing simultaneous differentiation between signals generated by the at least two labeled reporters associated with one or more of target cell depletion, activation of white blood cell subpopulations, and cytokine production by the white blood cells.