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A simple and rapid method for reproducible transient expression experiments to quickly identify the best conditions before the development of a stable CHO cell line is described, using adalimumab as a model antibody. Firstly, the signal peptide (SP) is optimized using different in silico programs, of which those having a D score of more than 0.85 will be selected. A list of useful SPs is provided. Next, guidelines for selecting the codon optimization algorithm is provided based on a CAI value of more than 0.95. After that, the codon optimized genes of the heavy chain (HC) and light chain (LC) of an antibody with different SPs are engineered into an expression vector and the two recombinant plasmids are co-transfected into a CHO cell line of interest. The antibody titer, specific productivity (Qp), and viable cell density (VCD) are then checked by ELISA and flow cytometry analysis. The best SP pairs can then be used for the development of stable CHO cell lines.
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http://dx.doi.org/10.1007/978-1-0716-4104-0_4 | DOI Listing |
Biol Pharm Bull
September 2025
Laboratory of Health Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, 6-3 Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Parthanatos is a noncanonical form of regulated cell death mediated by the overactivation of poly(ADP-ribose) polymerase-1, yet its regulatory mechanisms are not fully understood. To fully elucidate its regulatory mechanisms, it is necessary to establish useful research tools to investigate parthanatos. We have previously identified the human fibrosarcoma HT1080 cells as highly sensitive cells to parthanatos, and cefotaxime, a 3rd-generation cephem antibiotic, as the parthanatos inducer.
View Article and Find Full Text PDFToxicology
September 2025
Brown University, Department of Pathology and Laboratory Medicine, Providence, RI 02903, USA. Electronic address:
Mercury (Hg) is a global contaminant that is present in human diet as methylmercury (MeHg). Recent studies linked MeHg exposure with high risks of skin cancers. It is unknown whether MeHg is directly genotoxic in skin cells or able to enhance mutagenic effects of UV radiation.
View Article and Find Full Text PDFJ Biol Chem
September 2025
Department of Oral Disease Research, National Center for Geriatrics and Gerontology, 7-430 Moriokacho, Obu, Aichi, 474-8511, Japan; Department of dental hygiene, Ogaki women's college, 109-1 Nishinokawa-cho, Ogaki-city, Gifu, 503-8554, Japan. Electronic address:
Phagocytosis is mediated mainly by immune cells, such as macrophages, monocytes and neutrophils, that clear large pathogens including bacteria. The small GTP-binding protein Rab5 is crucial for both clathrin-dependent endocytosis and phagocytosis, but the role and mechanism of Rab5 activation during phagocytosis are poorly understood. Here we report that nitric oxide (NO), a novel regulator of Rab5, regulates phagocytosis through S-nitrosylation of Rab5.
View Article and Find Full Text PDFEcotoxicol Environ Saf
September 2025
College of Environment and Climate, Guangdong Key Laboratory of Environmental Pollution and Health, Jinan University, Guangzhou, Guangdong 510632, China.
Synthetic antioxidants and plasticizers have emerged as environmental contaminants to which prenatal exposure is widespread, raising concerns about adverse pregnancy outcomes. This study aimed to investigate the association between prenatal exposure to a mixture of synthetic antioxidants and plasticizers and the risk of spontaneous preterm birth (SPB), alongside underlying molecular responses. A nested case-control design was established, including 80 SPB cases and 170 matched healthy controls.
View Article and Find Full Text PDFJ Phys Chem B
September 2025
Department of Physics and Chemistry, DGIST, Daegu 42988, Republic of Korea.
Epidermal growth factor receptor (EGFR) dimerization plays a pivotal role in cellular signaling, influencing proliferation and disease progression, particularly in cancer. Despite extensive studies, the quantitative relationship between EGFR expression levels and dimerization efficiency remains incompletely understood. In this study, we investigated EGFR dimerization kinetics using ensemble-level biochemical assays and single-molecule tracking (SMT) in living cells.
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