Generation of high-producing CHO cell lines using cell picking system combined with fluorescent labeling of secreted products
Year: 2026
Type: article
Abstract: To facilitate the commercialization of protein therapeutics, the establishment of a mass production system is necessary for cost reduction and ensuring a stable supply. The common method for producing protein therapeutics is a mammalian cell-based expression system, predominantly utilizing Chinese hamster ovarian (CHO) cells. In the process of generating stable CHO cell lines expressing protein therapeutics, the most critical step is the selection of high producer cell lines. While limiting dilution has been a classical method for selecting candidate clones, it is time-consuming and lacks the ability to evaluate cell productivity. To address this issue, in this study, a novel selection method utilizing Cell Picking System was established and compared with limiting dilution in identifying high-producer clones. In this method, transfected CHO cells were loaded into a microchamber containing 20-µm-diameter microwells, allowing secreted proteins from individual cells to be detected using fluorescently labeled antibodies. The results of small-scale production test demonstrated that clones with higher productivity distributions were enriched using Cell Picking System. In addition, the highest clone achieved 2.34 g/L protein production, suggesting potential applicability for commercial supply with some modifications in cell culture conditions. This method supports high-throughput applications of protein therapeutics to the industrial scale and addresses the limitations of traditional selection methods.
Source: Scientific Reports
Authors: Hiroto Takeuchi, Naoya Maekawa, Tomohiro Okagawa, Yayoi Kameda, Chie Nakajima, Shiro Murata, Kazuhiko Ohashi, Yasuhiko Suzuki, Satoru Konnai
Institutions: Hokkaido University, Fasmac Co., Ltd. (Japan)
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,690
Awards: G128463445, G1666536545, G198540823, G2057278983, G2241976358, Analysis of the molecular mechanism for immunosuppression in malignant infectious diseases of livestock and the development of a novel control method, G2526894258, G2592339377, Mechanism of tumorigenesis in bovine leukemia initiated by chimeric transcripts of BLV-host genes, Development of advanced therapeutic technologies to address unmet medical needs for intractable diseases in animals, G3572227062, G3775553658, G3792724488, G385053884, Development of immunotherapy targeting immunoinhibitory receptors for control of bovine leukemia virus infection, G4507775186, Elucidation of the mechanism of tumorigenesis in bovine leukemia by comprehensive genomic analysis of proviral integration sites, G4732634665, G5081400964, G520310200, G5319852535, G5515750834, G5858130863, Development of novel control strategy for livestock malignant infectious diseases targeting immune disruptors, G595243434, G6235440555, G6672810681, Establishment of early diagnosis method based on elucidation of the molecular mechanism of bovine leukemia pathogenesis, G7313320092, G7475744167, G8407486487, G8691330753, G8726200593, G974363664
OpenAlex ID: W7171106184
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