蛋白质芯片技术应用于高通量单克隆抗体制备研究

Protein Array Technology Applied in High Throughput Monoclonal Antibody Generation

摘要:

针对在传统的单克隆抗体制备过程中进行特异性筛选时大量的人力消耗，建立了一种联合应用蛋白质芯片进行单克隆抗体制备的方法。用8种重组蛋白分别免疫BALB/c小鼠，在传统的细胞融合的基础上，将8种抗原免疫的杂交瘤阳性细胞混合后进行克隆化、蛋白质芯片筛选，阳性细胞有限稀释克隆化制备相关抗体。实验结果：混合克隆化共得到单克隆细胞175孔，经蛋白质芯片筛选出阳性孔119孔，选择针对单一抗原阳性的细胞连续2轮克隆化，8种重组蛋白各获得单克隆抗体细胞株1株。与经典的单克隆抗体制备相比，蛋白质芯片筛选与混合克隆化技术联合应用于单克隆抗体制备，1个筛选周期获得了8种重组蛋白的单克隆抗体细胞株，提高了单克隆抗体的制备效率，节省了在筛选中的抗原用量，提供了一种经济、快速、简便的方法。

Abstract:

To reduce the huge labor-cost in the screening in traditional monoclonal antibody generation, We established a new system for monoclonal antibody generation integrating with protein array. BALB/c mice were immunized by eight recombinant proteins respectively, and the positive hybridoma cells were obtained by cell fusion and ELISA screening. All the eight kinds of positive hybridoma cells were mixed, cloned, screened by protein array, and definite dilution cloned. Results: 175 single cell clones were obtained by complex cloning, and 119 of those were positive clones. Then 8 positive cell lines were generated by the following 2 rounds definite dilution cloning. By comparing with the traditional method, we got 8 monoclonal antibodies using the combined protein array screening and multiplex cloning method in 1 cycle, and fewer amounts of antigens were used. As a result, the combined protein array and multiplex cloning method could be used as an economical, rapid and simple tool applying in high throughput monoclonal antibody generation.