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81.
Matthias Rüdt Philipp Vormittag Nils Hillebrandt Jürgen Hubbuch 《Biotechnology and bioengineering》2019,116(6):1366-1379
Virus-like particles (VLPs) have shown great potential as biopharmaceuticals in the market and in clinics. Nonenveloped, in vivo assembled VLPs are typically disassembled and reassembled in vitro to improve particle stability, homogeneity, and immunogenicity. At the industrial scale, cross-flow filtration (CFF) is the method of choice for performing reassembly by diafiltration. Here, we developed an experimental CFF setup with an on-line measurement loop for the implementation of process analytical technology (PAT). The measurement loop included an ultraviolet and visible (UV/Vis) spectrometer as well as a light scattering photometer. These sensors allowed for monitoring protein concentration, protein tertiary structure, and protein quaternary structure. The experimental setup was tested with three Hepatitis B core Antigen (HBcAg) variants. With each variant, three reassembly processes were performed at different transmembrane pressures (TMPs). While light scattering provided information on the assembly progress, UV/Vis allowed for monitoring the protein concentration and the rate of VLP assembly based on the microenvironment of Tyrosine-132. VLP formation was verified by off-line dynamic light scattering (DLS) and transmission electron microscopy (TEM). Furthermore, the experimental results provided evidence of aggregate-related assembly inhibition and showed that off-line size-exclusion chromatography does not provide a complete picture of the particle content. Finally, a Partial-Least Squares (PLS) model was calibrated to predict VLP concentrations in the process solution. values of 0.947–0.984 were reached for the three HBcAg variants. In summary, the proposed experimental setup provides a powerful platform for developing and monitoring VLP reassembly steps by CFF. 相似文献
82.
Karthigayan Gunalan Xiaohong Gao Sally Shu Lin Yap Soak Kuan Lai Andrea Ravasio Sundar Ganesan Hoi Yeung Li Peter R. Preiser 《Cellular microbiology》2020,22(9)
Plasmodium falciparum responsible for the most virulent form of malaria invades human erythrocytes through multiple ligand‐receptor interactions. The P. falciparum reticulocyte binding protein homologues (PfRHs) are expressed at the apical end of merozoites and form interactions with distinct erythrocyte surface receptors that are important for invasion. Here using a range of monoclonal antibodies (mAbs) against different regions of PfRH1 we have investigated the role of PfRH processing during merozoite invasion. We show that PfRH1 gets differentially processed during merozoite maturation and invasion and provide evidence that the different PfRH1 processing products have distinct functions during invasion. Using in‐situ Proximity Ligation and FRET assays that allow probing of interactions at the nanometre level we show that a subset of PfRH1 products form close association with micronemal proteins Apical Membrane Antigen 1 (AMA1) in the moving junction suggesting a critical role in facilitating junction formation and active invasion. Our data provides evidence that time dependent processing of PfRH proteins is a mechanism by which the parasite is able to regulate distinct functional activities of these large processes. The identification of a specific close association with AMA1 in the junction now may also provide new avenues to target these interactions to prevent merozoite invasion. 相似文献
83.
84.
Microbial 2,3-butanediol production: a state-of-the-art review 总被引:7,自引:0,他引:7
2,3-Butanediol is a promising bulk chemical due to its extensive industry applications. The state-of-the-art nature of microbial 2,3-butanediol production is reviewed in this paper. Various strategies for efficient and economical microbial 2,3-butanediol production, including strain improvement, substrate alternation, and process development, are reviewed and compared with regard to their pros and cons. This review also summarizes value added derivatives of biologically produced 2,3-butanediol and different strategies for downstream processing. The future prospects of microbial 2,3-butanediol production are discussed in light of the current progress, challenges, and trends in this field. Guidelines for future studies are also proposed. 相似文献
85.
用免疫细胞化学和原位杂交技术探讨G、D细胞及胃泌素mRNA与肠化生的关系。标本来自胃镜活检的胃粘膜。结果显示,在与大肠化生区相邻的胃粘膜,G细胞突变消失,假幽门腺化生也缺乏G细胞,而淖肠化生仍保留少数G细胞;D细胞不仅见于小肠化生,而且也出现在假幽门腺化生以及某些大肠化生区。胃泌素mRNA仅限于G细胞分布区,未出现在大肠化生区和假幽门腺化生区,G细胞及胃泌素mRNA在大肠化生区的消失,可能由于局部杯状细胞分泌的硫酸粘蛋白改变了局部的微环境,从而影响了G细胞的分化与发育,至于假幽门腺化生区G细胞及胃泌素mRNA消失的原因还不清楚,应继续研究。 相似文献
86.
87.
免疫稳态的维持涉及多种细胞因子的基因表达调控,其中在转录后水平对mRNA稳定性的调控起重要作用。ARE(AU-rich element)位于mRNA 3'UTR(非编码区),富含AU碱基,某些RNA结合蛋白通过识别结合ARE影响mRNA的稳定性。本文综合最新研究,概述了TTP、HUR等RNA结合蛋白对mRNA稳定性的调节机制及其在信号通路中的作用。 相似文献
88.
研究戈壁地区单个灌丛及其下沙堆这一有机整体对周边土壤风蚀的抑制能力, 对加强相关地区的植被类型及其空间配置格局的防沙效应研究十分重要, 可为荒漠化监测的评价和制定科学的防治措施提供参考。该文利用数字图像处理技术, 获取吉兰泰盐湖北部戈壁上单个白刺(Nitraria tangutorum)灌丛沙堆和沙冬青(Ammopiptanthus mongolicus)灌丛沙堆周边地表不同土壤风蚀颗粒的百分含量; 并采用经典描述性统计及地统计学方法, 对各类土壤风蚀颗粒百分含量的水平空间异质性进行分析。结果表明: (1)灌丛基部和下风向是细物质积累区, 以灌丛堆为中心向外, <0.42 mm的细颗粒含量呈减少趋势; 而且细物质积累的最大值出现在白刺灌丛的迎风侧附近, 沙冬青样地则相反, 出现在灌丛的背风侧附近。在沙源物质有限的戈壁中, 白刺的防风固沙作用集中体现在灌丛附近, 其水平空间尺度范围不及沙冬青, 这亦是白刺样地粗粒化程度高于沙冬青样地的原因。(2)白刺和沙冬青灌丛附近地表中粒径>0.84 mm (不可蚀)、0.84-0.42 mm (半可蚀)及<0.42 mm (高度可蚀)颗粒的空间异质性尺度分别为17.80 m、66.63 m、8.41 m和9.82 m、15.33 m、14.91 m, 均超出了灌丛冠幅覆盖范围, 空间自相关部分比例C/(C0 + C)在63.40%-99.96%之间, 由此推断灌丛沙堆附近的风沙流特征是造成相应尺度内土壤颗粒空间异质性的主要因子。(3)高度可蚀颗粒的空间异质性尺度略大于灌丛平均间距(8.77 m包括灌丛半径), 从防止土壤风蚀来看, 这说明研究区内的建群种灌丛间存在一定程度的相互促进关系, 有利于该区植被的稳定与发展。 相似文献
89.
A retained intron in the 3′‐UTR of Calm3 mRNA mediates its Staufen2‐ and activity‐dependent localization to neuronal dendrites 下载免费PDF全文
90.
B. Kopmels J. Mariani N. Delhaye-Bouchaud F. Audibert D. Fradelizi E. E. Wollman 《Journal of neurochemistry》1992,58(1):192-199
We recently reported an abnormal production of interleukin-1 (IL-1) in peripheral macrophages of several neurological mutant mice that exhibit patterns of neuronal degeneration, especially in the cerebellum. After in vitro activation by lipopolysaccharide acid (LPS), these macrophages hyperexpress IL-1 beta mRNA and hyperproduce IL-1 protein in comparison with +/+ controls. In the present study, focused on the staggerer mutant mice, we investigate if this genetic dysregulation is specific for IL-1 beta or if it reflects a generalized hyperexcitability of these macrophages. The hyperexpression of IL-1 beta mRNA in sg/sg macrophages is present whatever the duration of LPS stimulation, even for periods as short as 15 min, although it reaches a maximum after 4 h of stimulation. The hyperinducibility of sg/sg macrophages is observed even when very low doses of LPS are used (0.01 microgram/ml) and reaches its maximum for 5 micrograms/ml LPS. Synthetic molecules (muramyl dipeptides), such as N-acetylmuramyl-L-alanyl-D-isoglutamine or murabutide, known as macrophage activators, are also efficient in revealing the cytokine hyperexpression in sg/sg macrophages. In addition, hyperexpression of two other cytokines, i.e., tumor necrosis factor-alpha and IL-1 alpha mRNAs, is also detected in LPS-stimulated macrophages of mutant mice. Finally, the effect of an inhibitor of protein synthesis, cycloheximide, is similar in +/+ and sg/sg macrophages. As a whole, these data lead us to conclude that the sg/sg macrophages are in a state of general hyperexcitability when compared with +/+ ones. 相似文献