SBIR Phase I: Antigen-Mediated Selection of Hybridomas

9860120 该SBIR I期项目旨在开发一种快速、自动化的方法，使用琼脂糖凝胶微囊化技术和流式细胞术从异质群体中选择杂交瘤细胞。将单个杂交瘤细胞包封到琼脂糖凝胶微滴（GMD）中，所述琼脂糖凝胶微滴具有锚定到琼脂糖基质的蛋白质抗原，允许蛋白质抗原充当捕获分子。与锚定蛋白抗原相互作用的杂交瘤分泌的抗体将用GMD内的荧光标记的抗IgG抗体检测，从而允许基于分泌的抗体对蛋白抗原的亲和力通过荧光激活细胞分选（FACS）分离杂交瘤。将对显示高度荧光的含有GMD的细胞进行分选、合并、培养，随后在选择试验的第二次通过中进行筛选。最终结果将是分泌针对所需蛋白抗原的高亲和力抗体的杂交瘤细胞。目前用于筛选杂交瘤细胞的方法是劳动密集型的、昂贵的且不可靠的。 开发一种快速、自动化的方法来分离特异性或高抗体产生的目的杂交瘤细胞，将在大规模生产用于治疗或人类体外诊断用途的许多基因工程蛋白质中得到广泛应用。

9860120 This SBIR Phase I project is aimed at developing a rapid, automated method to select hybridoma cells from a heterogeneous population using agarose gel microencapsulation technology and flow cytometry. Individual hybridoma cells will be encapsulated into agarose gel microdrops (GMDs) that have a protein antigen anchored to the agarose matrix, allowing the protein antigen to function as the capture molecule. Hybridoma secreted antibodies that interact with the anchored protein antigen will be detected with a fluorescently labeled anti-IgG antibody within the GMD, permitting isolation of hybridomas by fluorescently activated cell sorting (FACS) based on the affinity of the secreted antibodies for the protein antigen. GMD-containing displaying a high degree of fluorescence will be sorted, pooled, cultured, and subsequently screened in a second pass of the selection assay. The end result will be hybridoma cells secreting high affinity antibodies directed towards the desired protein antigen. Current methods for screening hybridoma cells are labor intensive, costly, and unreliable. Development of a rapid, automated method to isolate specific or high antibody producing hybridoma cells of interest would find broad application in the large scale production of many genetically engineered proteins for therapeutic or human in vitro diagnostic use.