Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 26, 6525 GA Nijmegen, Netherlands.
Division of Immunology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, Netherlands.
Sci Adv. 2019 Aug 28;5(8):eaaw1822. doi: 10.1126/sciadv.aaw1822. eCollection 2019 Aug.
Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete antibodies in the preferred format, species, and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab' fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemoenzymatic modification. We believe that this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research.
杂交瘤技术是开发新型抗体治疗药物和诊断试剂的重要手段。最近的临床前和临床研究强调了抗体同种型对于治疗效果的重要性。然而，由于恒定区的编码序列是固定的，因此在杂交瘤中调节抗体功能受到限制。在这里，我们展示了一种通用的 CRISPR/HDR 平台，可快速工程化恒定免疫球蛋白结构域，从而获得分泌所需形式、物种和同种型抗体的重组杂交瘤。利用该平台，我们获得了分泌 Fab'片段、同种型转换嵌合抗体和 Fc 沉默突变体的重组杂交瘤。这些抗体产品稳定，保留其抗原特异性，并在体外和体内显示其内在的 Fc 效应功能。此外，我们可以通过化学酶修饰将货物特异性地连接到这些抗体产品上。我们相信，这个通用的平台为整个科学界提供了便利，为临床前抗体研究提供了支持。
