应用流式细胞仪检测聚合物纳米胶束表面抗体偶联效率

Determination of antibody coupling efficiency on nanoscale polymer micelle surface by flow cytometry

目的探索应用流式细胞仪(flow cytometer,FCM)检测纳米级聚合物胶束(polymeric micelles,PMs)表面偶联抗体效率的方法。方法将DMPE-PEG-Maleimide、DPPE-PEG-Maleimide、DSPE-PEG-Maleimide、DOPE-PEG-Malei-mide、DLPE-PEG-Maleimide聚合物纳米胶束分别与Anti-GOLPH2抗体共同孵育,使其彼此偶联;再与SNU-423细胞共同孵育,采用FCM检测SNU-423细胞膜表面GOLPH2抗原的阳性表达率,并比较各类聚合物纳米胶束与AntiGOLPH2抗体的相对偶联效率。结果 SNU-423细胞表面GOLPH2抗原的阳性表达率为1.23%;在各类聚合物纳米胶束中,DLPE-PEG-Maleimide与Anti-GOLPH2抗体的相对偶联效率最高,且与抗体浓度呈正相关。结论应用FCM检测聚合物纳米胶束表面抗体偶联效率的方法,能在一定程度上反映抗体与聚合物纳米胶束的抗体偶联情况,其检测结果具有一定的参考价值。

Objective To develop a method for determination of antibody coupling efficiency on nanoscale polymer micelle(PM)surface by flow cytometry(FCM).Methods DMPE-PEG-Maleimide,DPPE-PEG-Maleimide,DSPE-PEGMaleimide,DOPE-PEG-Maleimide and DLPE-PEG-Maleimide nanoscale polymer micelles were co-incubated with AntiGOLPH2 antibody for coupling,then co-incubated with SNU-423 cells.The positive rate of GOLPH2 antigen on SNU-423 cell membrane surface was determined by FCM,and the relative coupling efficiencies of polymer nanomicelles with AntiGOLPH2 antibody were determined.Results The positive rate of GOLPH2 antigen on SNU-423 cell surface was 1.23%.The relative coupling efficiency of DLPE-PEG-Maleimide and Anti-GOLPH2 antibody was the highest in various nanoscale polymer micelles,which was positively correlated to the antibody concentration.Conclusion FCM for determination of antibody coupling efficiency on nanoscale polymer micelle surface reflected the coupling of antibody to nanoscale polymer micelle at a certain degree,of which the result was of a certain reference significance.