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Structural Determination of the Polycystin-2 Channel by Electron Cryo-Microscopy
Published in Jinghua Hu, Yong Yu, Polycystic Kidney Disease, 2019
Commonly used mammalian host cell lines include human embryonic kidney (HEK293) and Chinese hamster ovary (CHO) cell lines.84 The HEK293 cell line is derived from original human embryo kidney cells transformed with sheared fragments of human adenovirus 5 (Ad5) DNA integrated into chromosome 19.85 It is capable of producing transcriptionally incompetent (E1-deleted) human adenoviral vectors.86 Several modified HEK293 cell lines, into which additional virus DNA components are integrated, are available. For example, HEK293E cells have Epstein–Barr virus nuclear antigen-1 (HEK293-EBNA1, or 293E) expressed constitutively, which offers a threefold improvement in recombinant protein yield with expression vectors bearing the Epstein–Barr virus origin of replication.87
Impact of N-glycosylation on Fcγ receptor / IgG interactions: unravelling differences with an enhanced surface plasmon resonance biosensor assay based on coiled-coil interactions
Published in mAbs, 2019
Florian Cambay, Olivier Henry, Yves Durocher, Gregory De Crescenzo
The CHO-3E7 and the HEK293-6E cell lines, stably expressing a truncated Epstein-Barr virus Nuclear Antigen-1 (EBNA1), were grown in suspension in serum-free FreeStyleTM F17 medium (Invitrogen, cat# A13835–01) supplemented with 4 mM glutamine (Sigma-Aldrich, cat# G8540), 0.1% v/v Kolliphor® P 188 (Sigma-Aldrich, cat# K4894). The HEK293-6E medium was also supplemented with 25 μg/mL of G418-Sulfate (Wisent, cat# 400–130-IG). Cultures were maintained between 0.1 and 2 × 107 cells/mL in 125 mL Erlenmeyer ventilated flasks shaken at 120 rpm in a humidified incubator at 37°C with 5% CO2. Cell density and viability were determined using a Cedex Innovatis automated cell counter (Roche), relying on a counting method based on the trypan blue exclusion method.
Increased antibody levels to stage-specific Epstein–Barr virus antigens in systemic autoimmune diseases reveal a common pathology
Published in Scandinavian Journal of Clinical and Laboratory Investigation, 2019
Louise Sternbæk, Anette H. Draborg, Mark T. Østerlund, Line V. Iversen, Lone Troelsen, Elke Theander, Christoffer T. Nielsen, Søren Jacobsen, Gunnar Houen
One environmental risk factor is infection with EBV, one of eight known human herpesviruses and one of the most common viruses found in humans. EBV infects approximately 95–99% of the world’s population, mostly during childhood [5]. Most primary EBV infections are asymptomatic, and EBV causes a persistent latent infection in memory B cells. The most important signature protein expressed is Epstein–Barr virus nuclear antigen 1 (EBNA1). EBNA1 plays an essential role in the transcription and replication of the viral DNA during the cell cycle, as it is required for replication and maintenance of the episomal EBV genome [6]. Occasionally, the virus switches to a productive infective stage called lytic cycle, by expressing all its genes and starting production of EBV virions [7, 8]. In this process, the virus goes through successive stages of latency and reactivation, denoted latency 0, I, II, III, early lytic, late lytic, envelopment, and release (budding).
Suitability of transiently expressed antibodies for clinical studies: product quality consistency at different production scales
Published in mAbs, 2022
Sara Rodriguez-Conde, Sophie Inman, Viv Lindo, Leanne Amery, Alison Tang, Uche Okorji-Obike, Wenjuan Du, Berend-Jan Bosch, Paul J. Wichgers Schreur, Jeroen Kortekaas, Isabel Sola, Luis Enjuanes, Laura Kerry, Katharina Mahal, Martyn Hulley, Olalekan Daramola
CHO-G22 is a derivative of the CHOK1 cell line (ECACC No: 85051005). This cell line was engineered to enhance transient protein expression by co-expressing the Epstein-Barr virus nuclear antigen-1 (EBNA-1) and glutamine synthetase genes. Cells were maintained in AZ proprietary medium supplemented with methionine sulfoximine (MSX; Merck, product ref. #M5379-500) and hygromycin (Sigma, product ref. #10687010). Cultures in Erlenmeyer flasks or roller bottles were incubated at 140 rpm in a humidified orbital shaking incubator at 36.5°C and 5% CO2.