Oliviero Claudia, Hinz Steffen C, Grzeschik Julius, Hock Björn, Kolmar Harald, Hagens Gerrit
Institute of Life Technologies, Haute Ecole d'Ingénierie HES-SO Valais Wallis, Sion, Switzerland.
Ferring Biologics Innovation Centre, Epalinges, Switzerland.
Methods Mol Biol. 2023;2681:343-359. doi: 10.1007/978-1-0716-3279-6_19.
Integration of a gene of interest (GOI) into the genome of mammalian cells is the first step of cell line development campaigns for the production of biotherapeutics. Besides random integration methods, targeted gene integration approaches have emerged as promising tools over the last few years. In addition to reducing heterogeneity within a pool of recombinant transfectants, this process can also facilitate shorter timelines of the current cell line development process. Herein, we describe protocols for generating host cell lines carrying matrix attachment region (MAR)-rich landing pads (LPs), including BxB1 recombination sites. These LP-containing cell lines allow for site-specific and simultaneous integration of multiple GOIs. The resulting transgene-expressing stable recombinant clones can be used for the production of mono- or multispecific antibodies.
将目的基因（GOI）整合到哺乳动物细胞基因组中是生物治疗药物生产细胞系开发活动的第一步。除了随机整合方法外，靶向基因整合方法在过去几年中已成为有前景的工具。除了减少重组转染子库中的异质性外，该过程还可以缩短当前细胞系开发过程的时间线。在此，我们描述了生成携带富含基质附着区域（MAR）的着陆平台（LP）（包括BxB1重组位点）的宿主细胞系的方案。这些含LP的细胞系允许多个GOI进行位点特异性和同时整合。所得表达转基因的稳定重组克隆可用于生产单特异性或多特异性抗体。
