Due to the increasing economic and social relevance of biotherapeutics, their production processes are continually being reconsidered and reoptimized in an effort to secure higher product concentrations and qualities. Monitoring the productivity of cultured cells is therefore a critically important part of the cultivation process. Traditionally, this is achieved by determining the overall product titer by high performance liquid chromatography (HPLC), and then calculating the specific cell productivity based on this titer and an associated viable cell density. Unfortunately, this process is typically time‐consuming and laborious. In this study, the productivity of Chinese Hamster Ovary (CHO) cells expressing a monoclonal antibody was analyzed over the course of the cultivation process. In addition to calculating the specific cell productivity based on the traditional product titer determined by HPLC analysis, culture productivity of single cells was also analyzed via flow cytometry using a cold capture assay. The cold capture assay is a cell surface labelling technique described by Brezinsky et al., which allows for the visualization of a product on the surface of the producing cell. The cell productivity results obtained via HPLC and the results of cold capture assay remained in great accordance over the whole cultivation process. Accordingly, our study demonstrates that the cold capture assay offers an interesting, comparatively time‐effective, and potentially cheaper alternative for monitoring the productivity of a cell culture. 相似文献
Sustained pain relief following radon spa therapy in patients suffering from chronic painful diseases has been well described. But still, the underlying mechanisms are not fully understood. We conducted the prospective and explorative RAD-ON01 study which included 103 patients who suffered from chronic painful musculoskeletal disorders of the spine and/or joints and present here the data of the examination of pro- and anti-inflammatory cytokines in the serum of the patients before and at weeks 6, 12 and 30 after therapy. While TNFα, IL-1β, IFNγ, IL-1Ra and IL-10 were not altered, TGFβ was temporarily significantly (p = 0.013) elevated 6 weeks after therapy. Importantly, this elevation positively correlated with lowered pain sensitivity (r = 0.41). Further, the amount of IL-18 in the serum positively correlated with lowered pain sensitivity. Therefore, IL-18 can be considered as predictive marker for pain sensitivity of radon spa patients. We conclude that alterations in TGFβ and general IL-18 levels in serum have prognostic and predictive value in situations of lowered pain by exposure of patients to very low-doses of radiation as it is the case in radon spa.
The flowers constitute one of the main sites of accumulation of phenolic compounds in plants of the Hypericum genus. In addition to their important pharmacological activities, some metabolites found in species from the section Brathys and Trigynobrathys appear to have chemotaxonomic significance according to the literature. HPLC analyses were carried out to assess the pattern and accumulation of the dimeric phloroglucinols, benzophenones, benzopyrans, flavonoids and a phenolic acid in flowers of Hypericum species native to southern Brazil and Peruvian Páramos. Qualitative and quantitative differences are reported. Uliginosin B and hyperoside were the main components detected in all species and with maximum concentrations up to 0.188 % in H. caprifoliatum and 5.987 % in H. andinum, respectively. The content of japonicin A varied from 0.003 to 0.087 % in H. myrianthum, while the yield of hyperbrasilol B ranged from 0.006 % in H. laricifolium to 0.011 % in H. caprifoliatum. The major compounds in H. polyanthemum and H. carinatum were the benzopyrans 6-isobutyryl-5,7-dimethoxy-2,2-dimethyl-benzopyran (HP1 = 0.200 %), 7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethyl-benzopyran (HP2 = 0.225 %) and 5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethyl-benzopyran (HP3 = 0.327 %) and benzophenones cariphenone A (0.309 %) and cariphenone B (0.062 %), respectively. Maximum amounts of chlorogenic acid, isoquercitrin, quercitrin and guaijaverin were observed, respectively, in H. campestre (1.458 %), H. andinum (1.161 %), H. carinatum (0.231 %) and H. laricifolium (0.404 %). The results obtained support the taxonomic evidence of the dimeric phloroglucinol derivatives at the section level. 相似文献
Transparency is widespread in nature, ranging from transparent insect wings to ocular tissues that enable you to read this text, and transparent marine vertebrates. And yet, cells and tissue models in biology are usually strongly light scattering and optically opaque, precluding deep optical microscopy. Here we describe the directed evolution of cultured mammalian cells toward increased transparency. We find that mutations greatly diversify the optical phenotype of Chinese Hamster Ovary cells, a cultured mammalian cell line. Furthermore, only three rounds of high-throughput optical selection and competitive growth are required to yield fit cells with greatly improved transparency. Based on 15 monoclonal cell lines derived from this directed evolution experiment, we find that the evolved transparency frequently goes along with a reduction of nuclear granularity and physiological shifts in gene expression profiles. In the future this optical plasticity of mammalian cells may facilitate genetic clearance of living tissues for in vivo microscopy. 相似文献