4-Antibody AG, Basel, Switzerland.
NBE Therapeutics AG, Basel, Switzerland.
Methods. 2014 Jan 1;65(1):57-67. doi: 10.1016/j.ymeth.2013.09.003. Epub 2013 Sep 12.
Over the last nearly three decades in vitro display technologies have played an important role in the discovery and optimization of antibodies and other proteins for therapeutic applications. Here we describe the use of retroviral expression technology for the display of full-length IgG on B lineage cells in vitro with a hallmark of a tight and stable genotype to phenotype coupling. We describe the creation of a high-diversity (>1.0E09 different heavy- and light-chain combinations) cell displayed fully human antibody library from healthy donor-derived heavy- and light-chain gene libraries, and demonstrate the recovery of high affinity target-specific antibodies from this library by staining of cells with a labeled target antigen and their magnetic- and flow cytometry-based cell sorting. The present technology represents a further evolution in the discovery of full-length, fully human antibodies using mammalian display, and is termed Retrocyte Display® (Retroviral B lymphocyte Display).
在过去近三十年中,体外展示技术在抗体和其他治疗性蛋白质的发现和优化方面发挥了重要作用。在这里,我们描述了使用逆转录病毒表达技术在体外 B 细胞系上展示全长 IgG 的情况,其特点是基因型与表型紧密且稳定偶联。我们描述了从健康供体来源的重链和轻链基因文库中创建一个多样性高(>1.0E09 种不同的重链和轻链组合)的细胞展示全人源抗体文库,并通过用标记的靶抗原染色细胞,以及基于磁珠和流式细胞术的细胞分选,从该文库中回收高亲和力的靶特异性抗体。这项技术代表了使用哺乳动物展示发现全长全人抗体的进一步发展,被称为 Retrocyte Display®(逆转录病毒 B 淋巴细胞展示)。