A Cytometric Dual-Bead Array for Analyzing PML-RARalpha Fusion Protein
This study investigated a cytometric dual-bead array used to analyze PML-RARalpha fusion protein.The carboxylated and aminated polystyrene beads were prepared and barcoded with either high or low brightness using a fluorescein isothiocyanate (FITC) penetration method.Using anti-RARalpha antibody,homotype control antibody,barcoded beads and phycoerythrin (PE)-labeled antibody,the analytical capability of the cytometric dual-bead array was tested with two cell models containing a NB4 cell line.Fusion protein levels were normalized using a PE mean fluorescence intensity (MFI) ratio (PE MFI of high brightness beads / PE MFI of low brightness beads).In flow cytometry,the cytometric dual-bead array could analyze PML(L)- RARalpha protein with high specificity,and this assay possessed analytical sensitivity of at least 0.6%.A dilution experiment also demonstrated a concordant result between the logarithm of the PE MFI ratio and the logarithm of the NB4 cell concentration (R2 = 0.9936).When compared with a cytometric single-bead assay,this array exhibited a lower relative standard deviation (R.S.D.) and a lower relative error (R.E.).In conclusion,using the MFI ratio can attenuate data error in fusion protein cytometric analysis,and the cytometric dual-bead array presented here may be of use in the quantification of PML-RARalpha fusion protein.
cytometric dual-bead array fusion protein
Wen-Jun Lan Hai-Yan Wang Lei Yan Zhe-Li Wang Jun-Jie Ding
Institute of Biomedical Engineering Shandong Institute of Light Industry Jinan, Shandong Province, C Institute of Materia Medica Jinan Jinxiuchuan Pharmaceutical Factory Jinan, Shandong Province, China
国际会议
2013 ICME International Conference on Complex Medical Engineering(2013 ICME复合医学工程国际会议)
北京
英文
274-279
2013-05-25（万方平台首次上网日期，不代表论文的发表时间）