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Optimized production strategy of the major capsid protein HPV 16L1 non-assembly variant in E. coli
(2020)
The capsid of human papillomavirus (HPV) consists of two capsid proteins - the major capsid protein L1 and the minor capsid protein L2. Assembled virus-like particles, which only consist of L1 proteins, are successfully applied as prophylactic vaccines against HPV infections. The capsid subunits are L1-pentamers, which are also reported to protect efficiently against HPV infections in animals.
The recombinant production of L1 has been previously shown in E. coli, yeast, insect cells, plants and mammalian cell culture. Principally, in E. coli-based expression system L1 shows high expression yields but the protein is largely insoluble. In order to overcome this problem reported strategies address fusion proteins and overexpression of bacterial chaperones. However, an insufficient cleavage of the fusion proteins and removal of co-purified chaperones can hamper subsequent down streaming.
We report a significant improvement in the production of soluble L1-pentamers by combining (I) a fusion of a N-terminal SUMO-tag to L1, (II) the heterologous co-expression of the chaperon system GroEL/ES and (III) low expression temperature. The fusion construct was purified in a 2-step protein purification including efficient removal of GroEL/ES and complete removal of the N-terminal SUMO-tag. The expression strategy was transferred to process-controlled high-cell-density fermentation with defined media according to the guidelines of good manufacturing practice. The produced L1 protein is highly pure (>95%), free of DNA (260:280 = 0.5) and pentameric. The production strategy yielded 5.73 mg of purified L1-pentamers per gram dry biomass. The optimized strategy is a suitable alternative for high yield L1-pentamer production and purification as a cheaper process for vaccine production.
The wet-flue-gas desulfurization (FGD) process plays an important role in removing water-soluble flue-gas components such as sulphur dioxide (SO2) and oxidized mercury compounds. Under the reducing environment of the FGD, there is the possibility of re-emission of the already absorbed mercury (Hg) to the gas phase, which may be diminished by the utilization of specific additives. In this study, the effect of two different additives on Hg re-emission from the aqueous phase and Hg partitioning in gypsum and filtrate of a lab-scale wet-limestone FGD is investigated. Furthermore, the behaviour of additives in the presence of different halides is studied. The studied additives are TMT 15® as a sulphidic precipitating agent, which forms non-soluble mercury compounds, and activated lignite (AL) as a carbon-based sorbent, which adsorbs Hg compounds from the aqueous phase. TMT 15® has no significant effect on SO2 absorption; on the other hand, addition of AL improves the SO2-removal efficiency by up to 30%. Using both additives, Hg re-emission is suppressed in all the experimented cases except for AL in the absence of halides, in which Hg re-emission shows no change. Thus, the need to form nucleophilic oxidized mercury compounds in the slurry for the adsorption of oxidized mercury on AL can be concluded. Usage of both additives improves Hg retention in the slurry to different extents. It is shown that, for the additive-free slurries, the Hg-adsorption capacity of the solid fraction of the slurry is the limiting parameter. Moreover, the utilization of both additives results in a significant increase in the Hg concentration of solid fraction. The correlation between redox potential and partitioning of Hg in the slurry is presented by comparing the change in the redox potential of slurries when additives are used.
In a recent study, Gebauer et al. addressed a fundamental question regarding the effects of mind-body practices (MBPs) on the self. Does the practice of MBPs in accordance with traditional contemplative traditions quiet the ego or is the practice of MBPs associated with increased self-centrality, which breeds self-enhancement bias? Both hypotheses were investigated in two separate studies with a longitudinal design. Study 1 included 93 participants, who regularly practiced yoga, and study 2 contained 162 participants, who regularly practiced loving-kindness meditation. In both studies, trait questionnaires of self-centrality and self-enhancement were taken after the practice of yoga (over the course of 15 weeks) or meditation (over the course of 4 weeks). Findings from both studies showed that participants scored higher on measures of self-enhancement and self-centrality after practicing yoga and meditation as compared with not practicing yoga and meditation. Based on these findings, Gebauer et al. argued that MBPs such as yoga and meditation do not quiet the ego, but instead lead to self-enhancement bias through increased self-centrality. We have concerns about the far-reaching conclusions made by Gebauer et al. regarding the effects of MBPs on the self. The key concerns refer to the conceptualization of the quiet ego and to the assessment of the psychological constructs investigated in this study. Gebauer et al. addressed a timely and important research question, but their far-reaching interpretations should be reconsidered due to conceptual and methodological ambiguities.
Comparing multidimensional sensor data from vehicle fleets with methods of sequential data mining
(2020)
Reading and understanding large amounts of sensor data from vehicle test drives becomes more and more important. In order to test vehicle components or analyze exhaust emissions in real test drives, the sensor data obtained from these test drives have to be comparable. Otherwise components or exhaust emissions are tested and analyzed under false conditions. The sensor data obtained during test drives are highly multidimensional which makes it even more complicated to identify recurring patterns. We present a process model to compare different test drives according to their sensor data and so give an answer to the question whether or not test drives in different cities, locations and environments are representative to real driving scenarios. The algorithms we use focus on segmentation of the individual multivariate test drive data and on clustering of the segments according to different methods. We present several segmentation and cluster methods and compare which of them is best suited for comparing test drives. The segmentation method we identified as best suited is based on principal component analysis. As cluster methods we examine hierarchical, partitioning and density-based clustering in detail.
This study investigates how integrated reporting (IR) creates value for investors. It examines how providers of financial capital benefit from an improved firm information environment provided by IR. Specifically, this study investigates the effect of voluntary IR disclosure on analyst earnings forecast accuracy as well as on firm value. To do so, we use an international sample of 167 listed companies that voluntarily publish an integrated report. Our analysis shows no significant effect of a voluntary IR publication on analyst earnings forecast accuracy and no significant effect on firm value. We thus do not find evidence for the fulfillment of IR's promises regarding improved information environment and value creation of voluntary adopters. We conclude that such companies might already have a relatively high level of transparency leading to an absent additional effect of IR disclosure. Positive effects of IR appear to be more relevant in environments where IR is mandatory.
