Fu Shengyu, Zhao Qi
Cancer Centre, Institute of Translational Medicine, Faculty of Health Sciences, University of Macau, Taipa, Macau SAR 999078, China.
MoE Frontiers Science Center for Precision Oncology, University of Macau, Taipa, Macau SAR 999078, China.
Antibodies (Basel). 2022 Aug 16;11(3):53. doi: 10.3390/antib11030053.
Since they lack native soluble membrane antigens, the analysis and selection of antigen-specific antibodies are commonly performed on whole live cells. Here, we have developed a simple and convenient enzyme-linked immunosorbent assay (ELISA) based on cell membrane antigens. Soluble cell membrane proteins isolated from Raji cells were immobilized on the polystyrene microplate, which permitted the assessment of a therapeutic anti-CD22 monoclonal antibody. The experiments showed less variability in the intra-assay. Compared to the living cell ELISAs, the advantage of the assay is avoiding cell losses and high variation of optical density (OD) readings. We provide a quantitative and reproducible ELISA that can be potentially applied to the development of specific antibodies against cell surface antigens.
由于它们缺乏天然的可溶性膜抗原，抗原特异性抗体的分析和选择通常在完整的活细胞上进行。在此，我们开发了一种基于细胞膜抗原的简单便捷的酶联免疫吸附测定（ELISA）。从Raji细胞中分离出的可溶性细胞膜蛋白被固定在聚苯乙烯微孔板上，这使得能够评估一种治疗性抗CD22单克隆抗体。实验显示批内变异较小。与活细胞ELISA相比，该测定的优点是避免了细胞损失以及光密度（OD）读数的高变异性。我们提供了一种定量且可重复的ELISA，其有可能应用于针对细胞表面抗原的特异性抗体的开发。
