| This study addressed the preparation of polystyrene beads, the encoding of fluorescein isothiocyanate (FITC) and the analysis of multiplexed bead array for acute promyelocytic leukemia (APL). Polystyrene beads in this study were synthesized by styrene polymerization reaction. FITC dye was used to encode polystyrene beads as two bead populations in which the high brightness beads were coated with anti-PML antibody to capture PML-RARαfusion protein, and the low brightness beads were conjugated with homotype control antibody to perform the background control. Report antibody identified by PE-labeled antibody can recognise RAR alpha peptide epitope in fusion protein captured by the high-light beads. Thus PML-RARαfusion protein can be quantified via detecting PE fluorescence intensity using flow cytometer.We firstly studied the effect of reagent adding approach, reaction time, stabilizer and crosslinker dosage on microspheres diameter and coefficient of variation. Electron microscope assay exhibited that 5.4μm polystyrene beads with functional groups can be successfully polymerized, when a mixture containing 6 g stabilizer, 0.3 g crosslinker, a carboxylic acid and a amino compound was reacted for 12 hr.Next this paper studied the action of FITC concentration and reaction time to functional beads encoding. Flow cytometry appeared that the high brightness and low brightness beads, which constructed the duplexed beads array, can be successfully encoded by 0.4μL and 125μL FITC fluorescence solution after 0.5 hour incubation.APL duplexed beads array were also tested in this study. Flow cytometry showed results as following: a, PML-RARαfusion protein in NB4 cell line can be specially detected by the duplexed beads array. b, The value of array sensitivity was 0.63%. c, The coefficient of variation between different batch array was less than 10%. d, There was a clear relationship between this array and a single bead population assay.
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