Table II. Parameters of magnetic bead handler protocol for sandwich immunoassay.

Step	Plate content	Volume	Mixing parameters
			Time	Temperature	Speed
		μl
1) Transfer	Magnetic bead array	50
2) Analyte capture	Crude cell culture lysate	100	Overnight	8 °C	750 rpm
3) Wash + transfer	Wash buffer	100	20 s	RT	Slow
4) Wash + transfer	Wash buffer	100	20 s	RT	Slow
5) Detection antibody (5 cycles)	Detection antibody mixture	100	10 min, pause
			2 min	RT	Very slow
6) Wash + transfer	Wash buffer	100	20 s	RT	Slow
7) Wash + transfer	Wash buffer	100	20 s	RT	Slow
8) Detection reagent (4 cycles)	SAPE solution	100	10 min, pause
			2 min	RT	Very slow
9) Wash + transfer	Wash buffer	100	20 s	RT	Slow
10) Wash + transfer	Wash buffer	100	20 s	RT	Slow
11) Bead release to assay plate	Assay buffer	100	20 s	RT	Slow