Immunotechnology Group, Institute of Biochemistry and Biology, University of Potsdam, Potsdam, Germany.
Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany.
Sci Rep. 2020 Feb 3;10(1):1664. doi: 10.1038/s41598-020-58571-w.
The use of monoclonal antibodies is ubiquitous in science and biomedicine but the generation and validation process of antibodies is nevertheless complicated and time-consuming. To address these issues we developed a novel selective technology based on an artificial cell surface construct by which secreted antibodies were connected to the corresponding hybridoma cell when they possess the desired antigen-specificity. Further the system enables the selection of desired isotypes and the screening for potential cross-reactivities in the same context. For the design of the construct we combined the transmembrane domain of the EGF-receptor with a hemagglutinin epitope and a biotin acceptor peptide and performed a transposon-mediated transfection of myeloma cell lines. The stably transfected myeloma cell line was used for the generation of hybridoma cells and an antigen- and isotype-specific screening method was established. The system has been validated for globular protein antigens as well as for haptens and enables a fast and early stage selection and validation of monoclonal antibodies in one step.
单克隆抗体在科学和生物医学中被广泛应用，但抗体的产生和验证过程仍然复杂且耗时。为了解决这些问题，我们开发了一种新型的选择性技术，该技术基于人工细胞表面构建体，当分泌的抗体具有所需的抗原特异性时，它们会与相应的杂交瘤细胞连接。此外，该系统还能够在同一环境中选择所需的同种型并筛选潜在的交叉反应性。为了设计该构建体，我们将表皮生长因子受体的跨膜结构域与血凝素表位和生物素接受肽结合，并对骨髓瘤细胞系进行转座子介导的转染。稳定转染的骨髓瘤细胞系被用于生成杂交瘤细胞，并建立了一种抗原和同种型特异性筛选方法。该系统已针对球状蛋白抗原以及半抗原进行了验证，并能够在一步中快速、早期地选择和验证单克隆抗体。
