Koskela Essi V, Gonzalez Salcedo Alina, Piirainen Mari A, Iivonen Heidi A, Salminen Heidi, Frey Alexander D
Department of Bioproducts and Biosystems, Aalto University, Espoo, Finland.
Front Bioeng Biotechnol. 2020 Mar 31;8:255. doi: 10.3389/fbioe.2020.00255. eCollection 2020.
is a common platform for production of therapeutic proteins, but it is not intrinsically suited for the manufacturing of antibodies. Antibodies are naturally produced by plasma cells (PCs) and studies conducted on PC differentiation provide a comprehensive blueprint for the cellular transformations needed to create an antibody factory. In this study we mined transcriptomics data from PC differentiation to improve antibody secretion by . Through data exploration, we identified several new target genes. We tested the effects of 14 genetic modifications belonging to different cellular processes on protein production. Four of the tested genes resulted in improved antibody expression. The ER stress sensor increased the final titer by 1.8-fold and smaller effects were observed with , , and increasing antibody titers by 1. 6-, 1. 4-, and 1.4-fold. When testing combinations of these genes, the highest increases were observed when co-expressing with , or with and , resulting in 3.8- and 3.1-fold higher antibody titers. In contrast, strains expressing alone or in combination with the other genes produced similar or lower levels of recombinantly expressed endogenous yeast acid phosphatase compared to the controls. Using a genetic UPR responsive GFP reporter construct, we show that acts through constitutive activation of the unfolded protein response. Moreover, the positive effect of expression was transferable to other antibody molecules. We demonstrate how data exploration from an evolutionary distant, but highly specialized cell type can pinpoint new genetic targets and provide a novel concept for rationalized cell engineering.
是生产治疗性蛋白质的常见平台，但它本质上并不适合制造抗体。抗体由浆细胞（PCs）自然产生，对PC分化进行的研究为创建抗体工厂所需的细胞转化提供了全面蓝图。在本研究中，我们挖掘了PC分化的转录组学数据以提高通过……分泌抗体的能力。通过数据探索，我们鉴定了几个新的靶基因。我们测试了属于不同细胞过程的14种基因修饰对蛋白质生产的影响。其中4个测试基因导致抗体表达得到改善。内质网应激传感器使最终效价提高了1.8倍，观察到……、……和……对抗体效价的影响较小，分别提高了1.6倍、1.4倍和1.4倍。当测试这些基因的组合时，共表达……与……，或……与……和……时观察到最高的增加，导致抗体效价分别提高3.8倍和3.1倍。相比之下，单独表达……或与其他基因组合表达的菌株与对照相比，重组表达的内源性酵母酸性磷酸酶水平相似或更低。使用遗传的未折叠蛋白反应（UPR）响应性绿色荧光蛋白报告构建体，我们表明……通过组成性激活未折叠蛋白反应起作用。此外，……表达的积极作用可转移到其他抗体分子上。我们展示了如何从进化距离较远但高度特化的细胞类型的数据探索中确定新的遗传靶点，并为合理的细胞工程提供新的概念。
