Recombinant antibodies for antibody arrays

描述(由申请人提供)：我们建议开发一种快速、低成本和高通量的方法来产生具有改进特性的重组工程抗体，用于蛋白质组学和免疫分析应用。通过直接修饰产生抗体的杂交瘤细胞系中的抗体基因，将产生和表达重组Fab片段和嵌合抗体等工程化抗体构建物，从而消除基因克隆和转基因的需要。在这个第一阶段的SBIR中，我们将通过修改十(10)个杂交瘤细胞株来证明其可行性，这些杂交瘤细胞株产生的抗体用于MSD的商业免疫分析试剂盒。我们将证明，我们可以修改细胞系，使它们将其抗体表达为重组Fab片段或重组嵌合体，该嵌合体包含链接到蛋白质结构域的Fab片段，从而实现位点特异性标记。 使用已建立的试剂盒中的抗体将使直接比较新抗体构建体与原始全长抗体的检测性能变得直接。该计划的长期目标是利用嵌合结构的有益特性--较低的非特异性相互作用和通过位点特异性标记控制固定化的能力--从而能够开发用于蛋白质组应用的高密度阵列。这些改进的多重免疫分析方法将在研究和临床诊断应用中具有重要价值。

DESCRIPTION (provided by applicant): We propose to develop a rapid, low cost and high-throughput method for generating recombinant engineered antibodies with improved properties for proteomic and immunoassay applications. Engineered antibody constructs such as recombinant Fab fragments and chimeric antibodies will be generated and expressed by directly modifying the antibody genes in antibody-producing hybridoma cell lines, thereby eliminating the need for gene cloning and transfection. In this Phase I SBIR, we will demonstrate feasibility by modifying ten (10) hybridoma cell lines that produce antibodies used in MSD's commercial immunoassay kits. We will demonstrate that we can modify the cell lines so that they express their antibodies as recombinant Fab fragments or recombinant chimeras comprising Fab fragments linked to protein domains that enable site- specific labeling. The use of antibodies from established kits will make it straightforward to directly compare the assay performance of the new antibody constructs to the original full length antibodies. The long term goal of the program is to take advantage of beneficial properties of the chimeric constructs - lower non-specific interactions and the ability to control immobilization through site-specific labeling - to enable the development of high density arrays for proteomic applications. These improved multiplexed immunoassays will have significant value in both research and clinical diagnostics applications.