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71.
Walter Grabherr 《Plant Systematics and Evolution》1941,90(1):53-62
Ohne Zusammenfassung 相似文献
72.
Artur Schuller Monika Cserjan-Puschmann Christoph Köppl Reingard Grabherr Martin Wagenknecht Matteo Schiavinato Juliane C. Dohm Heinz Himmelbauer Gerald Striedner 《Biotechnology journal》2021,16(3):2000376
The production of recombinant proteins usually reduces cell fitness and the growth rate of producing cells. The growth disadvantage favors faster-growing non-producer mutants. Therefore, continuous bioprocessing is hardly feasible in Escherichia coli due to the high escape rate. The stability of E. coli expression systems under long-term production conditions and how metabolic load triggered by recombinant gene expression influences the characteristics of mutations are investigated. Iterated fed-batch-like microbioreactor cultivations are conducted under production conditions. The easy-to-produce green fluorescent protein (GFP) and a challenging antigen-binding fragment (Fab) are used as model proteins, and BL21(DE3) and BL21Q strains as expression hosts. In comparative whole-genome sequencing analyses, mutations that allowed cells to grow unhindered despite recombinant protein production are identified. A T7 RNA polymerase expression system is only conditionally suitable for long-term cultivation under production conditions. Mutations leading to non-producers occur in either the T7 RNA polymerase gene or the T7 promoter. The host RNA polymerase-based BL21Q expression system remains stable in the production of GFP in long-term cultivations. For the production of Fab, mutations in lacI of the BL21Q derivatives have positive effects on long-term stability. The results indicate that adaptive evolution carried out with genome-integrated E. coli expression systems in microtiter cultivations under industrial-relevant production conditions is an efficient strain development tool for production hosts. 相似文献
73.
74.
Anita Eberl Thomas Altendorfer-Kroath Denise Kollmann Thomas Birngruber Frank Sinner Reingard Raml Christoph Magnes 《Analytical biochemistry》2016
2H2O as nonradioactive, stable marker substance is commonly used in preclinical and clinical studies and the precise determination of 2H2O concentration in biological samples is crucial. However, aside from isotope ratio mass spectrometry (IRMS), only a very limited number of methods to accurately measure the 2H2O concentration in biological samples are routinely established until now. 相似文献
75.
Eduard Puente-Massaguer Paula Grau-Garcia Florian Strobl Reingard Grabherr Gerald Striedner Martí Lecina Francesc Gòdia 《Biotechnology journal》2021,16(4):2000391
Stable cell pools are receiving a renewed interest as a potential alternative system to clonal cell lines. The shorter development timelines and the capacity to achieve high product yields make them an interesting approach for recombinant protein production. In this study, stable High Five cell pools are assessed for the production of a simple protein, mCherry, and the more complex HIV-1 Gag-eGFP virus-like particles (VLPs). Random integration coupled to fluorescence-activated cell sorting (FACS) in suspension conditions is applied to accelerate the stable cell pool generation process and enrich it with high producer cells. This methodology is successfully transferred to a bioreactor for VLP production, resulting in a 2-fold increase in VLP yields with respect to shake flask cultures. In these conditions, maximum viable cell concentration improves by 1.5-fold, and by-product formation is significantly reduced. Remarkably, a global increase in the uptake of amino acids in the Gag-eGFP stable cell pool is observed when compared with parental High Five cells, reflecting the additional metabolic burden associated with VLP production. These results suggest that stable High Five cell pools are a robust and powerful approach to produce VLPs and other recombinant proteins, and put the basis for future studies aiming to scale up this system. 相似文献
76.
George Grabherr 《植被学杂志》1995,6(4):601-601
Book reviewed in this article: Rundel, P.W., Smith, A.P. & Meinzer, F.C. (eds.) 1994. Tropical Alpine Environments. Plant form and function. xi + 376 pp. Cambridge University Press, Cambridge. ISBN 0–521–42089-X. Price: GBP 65.-. 相似文献