China
Africa
Explore chapters and articles related to this topic
Cell Line Development
Published in Wei-Shou Hu, Cell Culture Bioprocess Engineering, 2020
Recombinant proteins are produced in mammalian cells through a transient expression of the gene of interest (GOI), or by establishing a cell line that produces the GOI product permanently (Panels 6.2–6.4). Transient expression is frequently used for the generation of small quantities of proteins (up to a few grams) for protein characterization or toxicity testing. First, cells are transfected with a high concentration of plasmids that include the GOI. A large number of plasmids are taken up by cells into the cytoplasm. A fraction of those plasmids then enter the nucleus, where the GOI is transcribed into mRNA. The mRNA molecules are exported back to the cytosol, translated into proteins, and secreted into the medium. Due to the large number of plasmids transfected and the strong promoter used, a large amount of product is produced. The plasmids in the cell are diluted as the cells divide and degraded over time. Thus, the transient production of the protein is usually limited to a few days. Nevertheless, the process is easily carried out (requiring only plasmid preparation) and is frequently used to produce a small quantity of proteins for characterization and testing.
Auto-antibodies as Biomarkers for Disease Diagnosis
Published in Raj Bawa, János Szebeni, Thomas J. Webster, Gerald F. Audette, Immune Aspects of Biopharmaceuticals and Nanomedicines, 2019
Angelika Lueking, Heike Göhler, Peter Schulz-Knappe
An alternative strategy for the production of protein microarrays uses DNA as template immobilized onto a surface combined with an in vitro transcription and translation step. These approaches called nucleic-acid programmable protein array or DNA array to protein array (DAPA) (Sibani and LaBaer, 2011; He and Taussig, 2001) have the advantages that efforts for protein production can be circumvented and toxic proteins may be expressed in vitro. However, multiple process steps such as plasmid preparation, spotting of the DNA, and in vitro transcription and translation reactions are error prone and lead to considerable inter- and intra-batch differences, which may negatively affect the statistical and bioinformatical analysis.
Synthesis of iron oxide nanoparticles for DNA purification
Published in Journal of Dispersion Science and Technology, 2021
László Vanyorek, Ágnes Maria Ilosvai, Emma Szőri-Dorogházi, Csaba Váradi, Ferenc Kristály, Ádám Prekob, Béla Fiser, Tamás Varga, Zoltán Kónya, Béla Viskolcz
In nucleic acid isolation and binding capacity tests a high copy number vector is worthy to use due to the high signal-to-noise ratio, therefore pBAD type vector DNA have been used in our experiments. For initial plasmid preparation Escherichia coli (E. coli) culture was grown from single colony picked from a freshly streaked plate. This single colony was used to inoculate 5 ml of Luria Bertani (LB) medium and then this starter culture was incubated for 12–16 h at 37 °C with vigorous shaking in a suitable incubator.