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A high‐throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation 下载免费PDF全文
Lam Raga Anggara Markely Lila Cheung Young Jun Choi Thomas Ryll Scott Estes Shashi Prajapati Iva Turyan Ruth Frenkel Zoran Sosic James Lambropoulos Lia Tescione Thomas Ryll Melissa Berman 《Biotechnology progress》2016,32(1):235-241
The serum half‐life, biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high‐throughput method for semi‐quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:235–241, 2016 相似文献
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Hammond Hammond Stacy Denise Viehmannova Iva Zamecnik Jiri Panis Bart Hlasna Cepkova Petra 《Plant Cell, Tissue and Organ Culture》2019,138(3):559-570
Plant Cell, Tissue and Organ Culture (PCTOC) - Slow-growth is a biotechnological tool for medium-term conservation of plant germplasm under in vitro conditions. In the present study, we assessed... 相似文献
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Dakic Tamara Jevdjovic Tanja Lakic Iva Djurasevic Sinisa F. Djordjevic Jelena Vujovic Predrag 《Neurochemical research》2019,44(2):388-399
Neurochemical Research - Our group previously reported that 6-h fasting increased both insulin II mRNA expression and insulin level in rat hypothalamus. Given that insulin effects on central... 相似文献
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Conformational diversity of single‐stranded DNA from bacterial repetitive extragenic palindromes: Implications for the DNA recognition elements of transposases 下载免费PDF全文
Tatsiana Charnavets Jaroslav Nunvar Iva Nečasová Jens Völker Kenneth J. Breslauer Bohdan Schneider 《Biopolymers》2015,103(10):585-596
Repetitive extragenic palindrome (REP)—associated tyrosine transposase enzymes (RAYTs) bind REP DNA domains and catalyze their cleavage. Genomic sequence analyses identify potential noncoding REP sequences associated with RAYT‐encoding genes. To probe the conformational space of potential RAYT DNA binding domains, we report here spectroscopic and calorimetric measurements that detect and partially characterize the solution conformational heterogeneity of REP oligonucleotides from six bacterial species. Our data reveal most of these REP oligonucleotides adopt multiple conformations, suggesting that RAYTs confront a landscape of potential DNA substrates in dynamic equilibrium that could be selected, enriched, and/or induced via differential binding. Thus, the transposase‐bound DNA motif may not be the predominant conformation of the isolated REP domain. Intriguingly, for several REPs, the circular dichroism spectra suggest guanine tetraplexes as potential alternative or additional RAYT recognition elements, an observation consistent with these REP domains being highly nonrandom, with tetraplex‐favoring 5′‐G and 3′‐C‐rich segments. In fact, the conformational heterogeneity of REP domains detected and reported here, including the formation of noncanonical DNA secondary structures, may reflect a general feature required for recognition by RAYT transposases. Based on our biophysical data, we propose guanine tetraplexes as an additional DNA recognition element for binding by RAYT transposase enzymes. © 2015 Wiley Periodicals, Inc. Biopolymers 103: 585–596, 2015. 相似文献
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Goudarzi M Banisch TU Mobin MB Maghelli N Tarbashevich K Strate I van den Berg J Blaser H Bandemer S Paluch E Bakkers J Tolić-Nørrelykke IM Raz E 《Developmental cell》2012,23(1):210-218
Single-cell migration is a key process in development, homeostasis, and disease. Nevertheless, the control over basic cellular mechanisms directing cells into motile behavior in?vivo is largely unknown. Here, we report on the identification of a minimal set of parameters the regulation of which confers proper morphology and cell motility. Zebrafish primordial germ cells rendered immotile by knockdown of Dead end, a negative regulator of miRNA function, were used as a platform for identifying processes restoring motility. We have defined myosin contractility, cell adhesion, and cortex properties as factors whose proper regulation is sufficient for restoring cell migration of this cell type. Tight control over the level of these cellular features, achieved through a balance between miRNA-430 function and the action of the RNA-binding protein Dead end, effectively transforms immotile primordial germ cells into polarized cells that actively migrate relative to cells in their environment. 相似文献
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Vlkova M Rohousova I Drahota J Stanneck D Kruedewagen EM Mencke N Otranto D Volf P 《PLoS neglected tropical diseases》2011,5(10):e1344