细胞膜表面标志CD系列抗体蛋白芯片制备及应用的研究

目的：本研究利用抗原抗体免疫反应的原理,研制一种可同时快速检测多种蛋白的蛋白芯片,用于细胞膜表面抗原的检测。方法：将抗细胞表面标志CD抗原的CD系列抗体蛋白固定在醛基玻片上形成微阵列,从样品细胞中抽提膜蛋白抗原并用FSE标记,然后与固定在芯片上的CD抗体阵列反应,扫描仪检测反应结果。结果：本研究选用HL－60,K562,NB4,Jurkat细胞株为代表,测定到细胞膜表面蛋白质CD抗原HL－60,K562,NB4,Jurkat细胞CD13均为阳性,HL－60,NB4,Jurkat细胞CD4阳性,NB4细胞CD38阳性,K562细胞CD7阳性。所得结果与免疫印迹及免疫组织化学验证等方法一致。结论：HL－60,K562,NB4,Jurkat细胞膜表面蛋白质CD抗原有差异。蛋白芯片方法具有的高信息通量,低成本,制备,制定快速简单的优点,展示了良好的应用前景。

Preparation and application of protein microarray for CD seriesantibodies

Objective Based on antibody antigen reaction to develop protein microarrays for high throughput, rapid detection of multi proteins on cellular membrane surface. Methods Four types of cell lines, HL 60, K562, NB4 and Jurkat were used in this study. The antibodies for CD series were spotted on the aldehyde slide to form microarrays. The cell membrane proteins were isolated and labeled with FSE fluorescence so that the fluorescence signals could be scanned with chip scanner and analyzed after the proteins were captured by immobilized antibodies. The results from protein microarrays were compared with those with Western blot and cell immunohistochemistry. Results With the protein microarrays, HL 60, Jurkat, NB4 and K562 cells were all proved positive to CD13, the former 3 cell lines also showed CD4 positive, and NB4 and K562 cells had positive reaction with CD38 and CD7 antibodies respectively. All of these were identical with the results by Western blot and cell immunohistochemistry. Conclusion The technique of protein microarray is a rapid, simple, low cost and high throughput approach for detecting the CD series on cell membrane.