376,773 research outputs found
CHO-S master cell line generation.
No full text(A) The H11 locus was cleaved, using CRISPR/Cas9, to encourage integration of the landing pad donor. A successful knock-in at the H11 locus generates a master cell line that has two PhiC31 attP sites, and that co-expresses three proteins via a PGK promoter-driven transcript. (B) Genotyping of the 5’-arm and 3’-arm in the CHO-S master cell line. PCR was performed with genomic DNA and specific primer pairs. 1: CHO-S parental cell line with 5’-arm primers; 2: CHO-S master cell line ("4–6") with 5’-arm primers; 3: CHO-S parental cell line with 3’-arm primers; 4: CHO-S master cell line ("4–6") with 3’-arm primers.</p
Characterization of VH411-S-Tag peptide conjugate binding to the CHO-hLDLR-EGFP cell line.
No full text(A) Representative confocal photomicrographs of CHO-hLDLR-EGFP and CHO-hTfR-EGFP cells (green) incubated 1 hr at 37°C with 10 μM of VH411-S-Tag detected post-fixation with an anti-S-Tag and A647-conjugated secondary antibody (cyan) and 10 μg/mL of DiI-LDL. Cell nuclei are labeled with Hoechst#33258 (blue) at 0.5 μg/mL. Co-labeling appears in blue-green/orange in the merged pictures. Note that VH411-S-Tag co-localizes with hLDLR and is internalized by CHO-hLDLR-EGFP cells. (B) Representative confocal photomicrographs of CHO-hLDLR-EGFP cells (green) incubated 1 hr at 37°C with a macromolecular complex (see scheme Fig 3B) resulting from the co-incubation and interaction of 10 μM VH411-S-Tag peptide or VH411Sc-S-Tag peptide with the anti-S-Tag antibody (1/200) and the Alexafluor 647-conjugated secondary antibody (1/800) (cyan). Cells were concomitantly exposed to 10 μg/mL of DiI-LDL (red). Cell nuclei are labeled with Hoechst#33258 (blue). Co-labeling appears in blue-green/orange in the merged pictures.</p
Antigenicity and immunogenicity of SARS-CoV S protein receptor-binding domain stably expressed in CHO cells
No full textThe receptor-binding domain (RBD) of SARS coronavirus (SARS-CoV) spike (S) protein contains multiple conformation-dependent epitopes that induce neutralizing antibody responses. Here we used CHO-K1 cells to establish a cell line for stable expression of a 193-mer (residues 318-510) RBD (RBD193-CHO) and determined its antigenicity and immunogenicity. We found that RBD193-CHO reacted strongly with a panel of six monoclonal antibodies recognizing various conformational and linear epitopes in RBD, suggesting that this recombinant protein maintains intact conformation and good antigenicity. Immunization of mice with RBD193-CHO resulted in induction of high titers of RBD-specific neutralizing antibodies and potent IL-4-expressing T cell responses. RBD193-CHO induced immunity that protected a majority of the vaccinated mice from SARS-CoV challenge. These results suggest that the recombinant RBD produced in an established stable cell line maintains strong immunogenicity with high potential for use as an effective and economic subunit SARS vaccine. © 2009 Elsevier Inc. All rights reserved.link_to_OA_fulltex
Marriner S. Eccles correspondence related to Eccles quotations [07]
No full textCorrespondence from 1965 through 1967 between Marriner S. Eccles and friends, associates, and publishers who sent him published citations of and references to Mr. Eccles. Correspondents included economics author Irving S. Michelman; Hugh S. Norton, professor of economics at the University of South Carolina; and economist Eliot Janeway
A 219-mer CHO-expressing receptor-binding domain of SARS-CoV S protein induces potent immune responses and protective immunity
No full textDevelopment of vaccines is essential for the prevention of future recurrences of severe acute respiratory syndrome (SARS), caused by the SARS coronavirus (SARS-CoV). The spike (S) protein, especially receptor-binding domain (RBD) of SARS-CoV, plays important roles in the prevention of SARS infection, and is thus an important component in SARS vaccine development. In this study, we expressed a 219-mer (residues 318-536) RBD protein in Chinese hamster ovary (CHO)-K1 cells (RBD219-CHO), and tested its immune responses and protective immunity in a mouse model. The results showed that this recombinant protein was correctly folded, being able to maintain intact conformation and authentic antigenicity. It could induce strong humoral and cellular immune responses and high titers of neutralizing antibodies in the vaccinated mice. RBD219-CHO protein elicited potent protective immunity that protected all vaccinated mice from SARS-CoV challenge. These results suggest that the recombinant RBD219-CHO protein has great potential for the development of an effective and safe SARS subunit vaccine. © Copyright 2010, Mary Ann Liebert, Inc..published_or_final_versio
CHO microRNA engineering is growing up : recent successes and future challenges
No full textmicroRNAs with their ability to regulate complex pathways that control cellular behavior and phenotype have been proposed as potential targets for cell engineering in the context of optimization of biopharmaceutical production cell lines, specifically of Chinese Hamster Ovary cells. However, until recently, research was limited by a lack of genomic sequence information on this industrially important cell line. With the publication of the genomic sequence and other relevant data sets for CHO cells since 2011, the doors have been opened for an improved understanding of CHO cell physiology and for the development of the necessary tools for novel engineering strategies. In the present review we discuss both knowledge on the regulatory mechanisms of microRNAs obtained from other biological models and proof of concepts already performed on CHO cells, thus providing an outlook of potential applications of microRNA engineering in production cell lines
Rapid emergence of a viral resistant mutant in WHV chronically infected woodchucks treated with lamivudine and a pre-S/S CHO-derived hepatitis B virus vaccine
No full textTo determine whether the addition of a pre-S/S human vaccine increases the antiviral activity of lamivudine, four woodchucks were treated with a daily dose of 100 mg/kg lamivudine and four 50 microg doses of CHO-derived pre-S/S human vaccine. WHV DNA titres decreased up to two logarithms in three woodchucks. At week 4, in three of the animals, the sequence analysis showed a predominant strain containing a nucleotide change from A to T at position 1696 of domain B of the WHV DNA polymerase. Vaccination did not further suppress WHV DNA, despite anti-HBs production in three animals. The woodchuck remains a useful model for characterising the biology and kinetics of the emergence of drug-resistant variants and could be used for pre-clinical studies of combinations of new antiviral drugs
Diffusive author(s), cohesive author: Analysis of S/N (1994)
No full textThis study indicates the ways in which various aspects of the author(s) are brought forth in Dumb type’s performance art, the S/N production. Previous research has suggested a non-hierarchical organization of Dumb type and the absence of a “privileged author” in Dumb type’s collaborative work, S/N. However, the results that I have investigated from member’s interviews on the creative process of S/N along with my analysis of the recorded images of S/N, indicate a different aspect of the author(s). First, S/N was created through, so to speak, the collective ideas of the members of Dumb type. Further, S/N has at least nine quotations from previous performances, installations, and printed writings, besides the work-in-progress technique. Explicating one of the “author functions” as given by Michel Foucault, each text has plural subjects of the author. However, it has been revealed from members’ interviews that Teiji Furuhashi had a decision-making role in selecting the members’ ideas within the performance. Since then, S/N has had plural subjects of creation; however, Furuhashi is one of the subjects of creation along with the “privileged author.” S/N has plural authors (diffusive authors) yet at the same time, it has a “privileged author,” Teiji Furuhashi (cohesive author)
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