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. 2023 Sep 20;13:15620. doi: 10.1038/s41598-023-42925-1

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Characterization of the dual landing pads-master cell (dMCL). (A) Southern blot analysis of the number of integration sites of the landing pad 2 in dMCL. Genomic DNA of dMCL was digested with either XbaI, MfeI or NdeI, and resolved in 0.7% agarose gel. Membrane was hybridized with DIG-dUTP labeled probes targeting the NEO gene. One DNA band detected in each lane indicates single integration site of the landing pad 2. The expression cassette in landing pad 2 contained only one MfeI restriction site and none of XbaI and NdeI restriction sites. The dMCL were transfected with one or both of targeting vectors as shown in Fig. 1B to characterize expression capacities of each landing pad. (B) The stable cell pools generated by transfecting both targeting vectors were subjected to flow cytometry analysis. (C) Specific productivity (qP) of producer cell lines expressing mAbs from single landing pads and both landing pads during seven-day fed batch culture.