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生成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。通过在产生抗体的杂交瘤细胞系中直接修饰抗体基因，可以产生和表达重组Fab片段和嵌合抗体等工程抗体构建物，从而消除了基因克隆和转染的需要。在这个I期SBIR中，我们将通过修饰10（10）个杂杂瘤细胞系来证明其可行性，这些杂杂瘤细胞系可以产生用于默沙华公司商业免疫测定试剂盒的抗体。我们将证明，我们可以修饰细胞系，使它们以重组Fab片段或重组嵌合体的形式表达抗体，重组嵌合体包括连接到能够进行位点特异性标记的蛋白质结构域的Fab片段。

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.