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- Title
A novel aptamer beacon for rapid screening of recombinant cells and in vivo monitoring of recombinant proteins.
- Authors
Zheng, Wei-Wei; Yang, Li-Ting; Zhou, Hang; Zhang, Zhi-Yue; Wang, Xin-Yi; Wu, Jing-Yi; Lu, Xin-Chen; Chen, Jing; Duan, Cai-Wen; Chen, Kai-Ming
- Abstract
Recombinant protein drugs, which are typically produced by mammalian host cells, have been approved for the treatment of a range of diseases. Accordingly, systems for selecting recombinant cell lines with efficient protein expression and for testing the content of recombinant proteins in vivo are crucial to the large-scale production and application of protein-based therapeutic drugs. In this study, we designed three aptamer beacons to detect His-tag, a common label of recombinant proteins. We found that all three beacons could specifically and quantitatively measure the His-tagged recombinant proteins with a short reaction time. Among these three beacons, the 6H5-MU beacon had the highest sensitivity for His polypeptides with a detection limit of 250 ng/mL and the shortest detection time within 1 min. Furthermore, we established a rapid and highly effective recombinant cell line construction system, which could obtain monoclonal cell lines with high yields of target proteins within 21 days, by combining 6H5-MU with pSB, a novel plasmid composed of a Sleeping Beauty transposase and a transposon. Finally, 6H5-MU also discriminately tested the serum concentration of His-tagged recombinant proteins in vivo, with consistent results compared to enzyme-linked immunosorbent assay (ELISA). We thus established a rapid and high-throughput method for generating recombinant cell lines and in vivo monitoring of recombinant protein levels, thereby providing a new platform for the development and preparation of recombinant protein drugs. Key points: • The 6H5-MU aptamer beacon rapidly and accurately binds to His-tagged recombinant proteins. • A system for rapid and high-throughput generation of recombinant cell lines is established using 6H5-MU and pSB. • 6H5-MU allows in vivo monitoring of recombinant protein levels.
- Subjects
RECOMBINANT proteins; APTAMERS; TRANSPOSONS; ENZYME-linked immunosorbent assay; RECOMBINANT drugs; CELL lines; PROTEIN drugs
- Publication
Applied Microbiology & Biotechnology, 2023, Vol 107, Issue 2/3, p553
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-022-12331-0