Table 2.

Successive steps involved for the calibration and application of the prototype “IgHκ calibrator beads”.

Calibrator beads	QIFIkit (commercial)	Midi-QIFIkit (prototype)	Mini-QIFIkit (prototype)	µ-QIFIkit (prototype)	“IgHκ beads” (protoype)	“IgHκ beads” (prototype)
Diameter	10 µm	3 µm	3 µm	1 µm	1 µm	1 µm
Coated analyte	Mouse IgG1	Biotin-IgG1 (mouse)	Biotin-IgG1 (mouse)	Biotin-IgG1 (mouse)	REAfinity* CD151-biotin (human IgG-k)	Trastuzumab on liposomes (human IgG-k)
Coating method	Proprietary	Magnetic SA-beads	Magnetic SA-beads	Magnetic SA-beads	Magnetic SA-beads	Covalent coupling
Calibration FCM instrument	FC500 and/or Gallios	Gallios	CytoFLEX-S	CytoFLEX-S	CytoFLEX-S	CytoFLEX-S
Staining method	QIFI (indirect IF with unlabeled mouse IgGs)	QIFI (indirect IF with unlabeled mouse IgGs)	QIFI (indirect IF with unlabeled mouse IgGs)	QIFI (indirect IF with unlabeled mouse IgGs)	QIFI (indirect IF with unlabeled mouse IgGs)	Direct IF with L1C1-PE (anti-human kappa)
Laser height at interrogation point	= or >10 µm, (depending on instrument)	10 µm	5 µm	5 µm	5 µm	5 µm
Excitation λ	488 nm	488 nm	561 nm	561 nm	561 nm	561 nm
Calibration channel	FITC	FITC	PE	PE	PE	PE
Calibration curve(s)	N.A.	Fig. S1a	Fig. S1b	Fig. S1c	Fig. 1	Fig. 2