Table II Parameters of magnetic bead handler protocol for sandwich immunoassay
StepPlate contentVolumeMixing parameters
TimeTemperatureSpeed
μl
1) TransferMagnetic bead array50
2) Analyte captureCrude cell culture lysate100Overnight8 °C750 rpm
3) Wash + transferWash buffer10020 sRTSlow
4) Wash + transferWash buffer10020 sRTSlow
5) Detection antibody (5 cycles)Detection antibody mixture10010 min, pause
2 minRTVery slow
6) Wash + transferWash buffer10020 sRTSlow
7) Wash + transferWash buffer10020 sRTSlow
8) Detection reagent (4 cycles)SAPE solution10010 min, pause
2 minRTVery slow
9) Wash + transferWash buffer10020 sRTSlow
10) Wash + transferWash buffer10020 sRTSlow
11) Bead release to assay plateAssay buffer10020 sRTSlow