抗癌胚抗原抗体的交联浓度对液相芯片荧光检测信号的影响

Effect of coupled CEA antibody concentration on liquichip fluorescence detection signal

目的探讨不同交联浓度的抗癌胚抗原（CEA）单克隆抗体对液相芯片荧光检测信号的影响，明确CEA检测中最适抗体交联浓度。方法分别以不同交联浓度的标记有生物素的抗-CEA单抗与Luminex公司生产的44号荧光微球交联，交联完成后在每个反应孔中加入交联有不同浓度抗体的荧光微球各5000个，每个浓度加8孔，在每个抗体交联浓度下对应的检测孔中分别加入藻红蛋白（PE）标记的亲和素，经过15min的反应后用Luminex-100荧光检测仪检测荧光强度。结果CEA抗体的最适交联浓度为100μg／ml。结论在液相芯片的蛋白检测中CEA抗体有其最适的交联浓度。

Objective To explore the effect of different concentrations of coupled CEA antibody on liquichip fluorescence detection signal,and ascertain the optimal coupled CEA antibody concentration during CEA detection. Methods Anti-CEA monoclonal antibodies labelled with biotin at different concentrations （25, 50, 100, 150, 200μg/ml） were coupled to No. 44 fluorescent beads produced by Luminex Company. Each of 8 wells was added 5 000 beads coupled with the same concentration of antibody respectively. Different concentrations of avidin labeled with phycoerythrin （PE） were added the corresponding wells （0. 08, 0. 16, 0.32, 0.64, 2.56, 12.80, 64.00, 128.00 μg/ml corresponding to 25, 50, 100, 150,200 μg/ml of antibodies respectively）. After incubation for 15 min under 37℃ ,Luminex-100 was used to measure fluorescence intensity. Results The optimal coupled anti-CEA antibody concentration was 100 μg/ml. Conclusion There exists the optimal coupled CEA antibody concentration during liquiehip protein detection.