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391.
Salina Louie Amy Heidersbach Noelia Blanco Benjamin Haley Christopher M. Rose Peter S. Liu Mandy Yim Danming Tang Cynthia Lam Wendy N. Sandoval David Shaw Brad Snedecor Shahram Misaghi 《Biotechnology progress》2020,36(3):e2951
Chinese hamster ovary (CHO) cells have been adapted to grow in serum-free media and in suspension culture to facilitate manufacturing needs. Some CHO cell lines, however, tend to form cell aggregates while being cultured in suspension. This can result in reduced viability and capacity for single cell cloning (SCC) via limiting dilution, and process steps to mitigate cell aggregate formation, for example, addition of anti-cell-aggregation agents. In this study, we have identified endothelial intercellular cell adhesion molecule 1 (ICAM-1) as a key protein promoting cell aggregate formation in a production competent CHO cell line, which is prone to cell aggregate formation. Knocking out (KO) the ICAM-1 gene significantly decreased cell aggregate formation in the culture media without anti-cell-aggregation reagent. This trait can simplify the process of transfection, selection, automated clone isolation, and so on. Evaluation in standard cell line development of ICAM-1 KO and wild-type CHO hosts did not reveal any noticeable impacts on titer or product quality. Furthermore, analysis of a derived nonaggregating cell line showed significant reductions in expression of cell adhesion proteins. Overall, our data suggest that deletion of ICAM-1 and perhaps other cell adhesion proteins can reduce cell aggregate formation and improve clonality assurance during SCC. 相似文献
392.
Danming Tang Cynthia Lam Niels Bauer Simon Auslaender Brad Snedecor Michael W. Laird Shahram Misaghi 《Biotechnology progress》2022,38(2):e3228
In the field of therapeutic protein production, process intensification strategies entailing higher starting cell seeding densities, can potentially increase culture productivity, lower cost of goods and improve facility utilization. However, increased cell densities often trigger apoptotic cell death at the end of the cell culture process and thus reduce total viable cell count. Apoptosis-resistant Chinese hamster ovary cell lines may offer the possibility to diminish this undesired outcome of the intensified production process. In this study, we have generated and tested Bax/Bak double-knock-out (DKO) apoptosis resistant hosts to express standard and bispecific antibodies, as well as complex molecules in intensified production processes both as pools and single cell clones, and at different scales. In all cases, therapeutic proteins expressed from clones or pools generated from the Bax/Bak DKO hosts showed not only better viability but also enabled extended productivity in the later stages of the 14-day intensified production process. The product qualities of the produced molecules were comparable between Bax/Bak DKO and wild type cells. Overall, we showed that Bax/Bak DKO apoptosis-resistant host cell lines significantly improve viability and volumetric productivity of the intensified production cultures without altering product qualities. 相似文献
393.
Amir Biranvand Oldřich Nedvĕd Samira Karimi Hassanali Vahedi Shahram Hesami Hossein Lotfalizadeh 《法国昆虫学会纪事》2020,56(2):106-114
Summary Over the past few decades, many records of the parasitoids parasitizing Coccinellidae have been reported from Iran. Some of them, presented in hard to access conference proceedings, theses, older books and journal articles, were then inaccurately quoted in other papers. Furthermore, some other records were subsequently considered misidentifications and the corrected identity of the parasitoids was proposed in later published papers. Due to the accumulation of that misleading information, we decided to gather and critically evaluate all available Iranian data on the parasitoids of Coccinellidae. To the review presented here we also added new data on the parasitism of Coccinella septempunctata L. by a braconid Dinocampus coccinellae (Schrank) in the Lorestan province and on the parasitism of Hyperaspis pseudopustulata Mulsant by an encyrtid Homalotylus flaminius (Dalman) in the Kermanshah province. Hyperaspis pseudopustulata is a new host for H. flaminius, and is also a species new to the fauna of Iran. Altogether, our review revealed one species of dipteran and 16 species of hymenopteran parasitoids associated with Coccinellidae in Iran. The identity of some of those species, however, seems uncertain and should be re-examined. 相似文献