Tu Bailin, Tieman Bryan, Moore Jeffrey, Pan You, Muerhoff A Scott
Abbott Diagnostics Division, Department of Biologics Discovery and Design, Abbott Laboratories , Abbott Park, Illinois.
Monoclon Antib Immunodiagn Immunother. 2017 Jun;36(3):113-118. doi: 10.1089/mab.2017.0009. Epub 2017 May 30.
Monoclonal antibodies are widely used as the capture and detection reagents in diagnostic immunoassays. In the past, myeloma fusion partners expressing endogenous heavy and/or light chains were often used to generate hybridoma cell lines. As a result, mixed populations of antibodies were produced that can cause inaccurate test results, poor antibody stability, and significant lot-to-lot variability. We describe one such scenario where the P3U1 (P3X63Ag8U.1) myeloma fusion partner was used in the generation of a hybridoma producing protein induced vitamin K absence/antagonist-II (PIVKA II) antibody. The hybridoma produces three subpopulations of immunoglobulin as determined by ion exchange (IEx) chromatography that exhibit varying degrees of immunoreactivity (0%, 50%, or 100%) to the target antigen as determined by Surface Plasmon Resonance. To produce an antibody with the highest possible sensitivity and specificity, the antigen-specific heavy and light chain variable domains (VH and VL) were cloned from the hybridoma and tethered to murine IgG1 and kappa scaffolds. The resulting recombinant antibody was expressed in Chinese hamster ovary cells and is compatible for use in a diagnostic immunoassay.
单克隆抗体被广泛用作诊断免疫分析中的捕获和检测试剂。过去，表达内源性重链和/或轻链的骨髓瘤融合伙伴常被用于生成杂交瘤细胞系。结果，产生了混合的抗体群体，可能导致检测结果不准确、抗体稳定性差以及批次间差异显著。我们描述了一种这样的情况，即P3U1（P3X63Ag8U.1）骨髓瘤融合伙伴被用于生成一种产生蛋白质诱导维生素K缺乏/拮抗剂-II（PIVKA II）抗体的杂交瘤。通过离子交换（IEx）色谱法测定，该杂交瘤产生三种免疫球蛋白亚群，通过表面等离子体共振测定，它们对靶抗原表现出不同程度的免疫反应性（0%、50%或100%）。为了产生具有尽可能高的灵敏度和特异性的抗体，从杂交瘤中克隆了抗原特异性重链和轻链可变区（VH和VL），并将其连接到小鼠IgG1和κ支架上。所得的重组抗体在中国仓鼠卵巢细胞中表达，适用于诊断免疫分析。
