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31.
Nathan A. Lacher Rachel K. Roberts Yan He Holly Cargill Katrina M. Kearns Heidi Holovics Margaret N. Ruesch 《Journal of separation science》2010,33(2):218-227
Capillary gel electrophoresis (CGE) methods with UV detection were developed for reduced and non‐reduced mAb analysis. These methods can be used to evaluate mAb purity, offering more reproducible quantitation compared with that of traditional SDS‐PAGE methods. These CGE methods have been utilized as platform technology for bioprocess development, formulation development, mAb characterization, drug substance/drug product release testing as well as a required methodology for stability testing. We have found these CGE methods to be applicable across a platform of mAbs in preclinical and clinical development, with the majority of mAbs requiring no modification to the method conditions. This methodology has been ICH validated and transferred to several supporting organizations. The data presented herein describes the development of CGE methodology, platform application to mAb purity analysis, ICH validation, reliability metrics, and considerations on technology enhancement for improved performance and throughput. 相似文献
32.
Nathan A. Lacher Qian Wang Rachel K. Roberts Heidi J. Holovics Serdar Aykent Michael R. Schlittler Melissa R. Thompson Charles W. Demarest 《Electrophoresis》2010,31(3):448-458
A CGE method for monitoring the disulfide isomer distribution characteristic of IgG2 MAbs is presented. Disulfide heterogeneity of MAbs has been studied using various chromatographic and electrophoretic methods. Although CGE operates using a different selectivity mechanism from that of sorption chromatographic techniques, similar trends are present in the data, which allow the CGE method to be used as a complementary method for studying disulfide isomer distribution. This article focuses on the optimization of a capillary‐based gel electrophoresis method that can be used to support antibody development including bioprocess optimization, antibody characterization, release, and formulation stability assessment. 相似文献
33.
Stephan Schulz Andreas Kuczkowski Heidi Saxell 《Journal of organometallic chemistry》2010,695(21):2281-2283
The Lewis acid-base adduct t-Bu3Al-SbMe3 (1), which was synthesized by reaction of equimolar amounts of t-Bu3Al and trimethylstibine SbMe3, was characterized by multinuclear NMR (1H, 13C) spectroscopy, elemental analyses as well as by single crystal X-ray diffraction. 相似文献
34.
Liesbeth Tytgat Matthias R. Kollert Lana Van Damme Hugo Thienpont Heidi Ottevaere Georg N. Duda Sven Geissler Peter Dubruel Sandra Van Vlierberghe Taimoor H. Qazi 《Macromolecular bioscience》2020,20(4)
Adipose tissue engineering aims to provide solutions to patients who require tissue reconstruction following mastectomies or other soft tissue trauma. Mesenchymal stromal cells (MSCs) robustly differentiate into the adipogenic lineage and are attractive candidates for adipose tissue engineering. This work investigates whether pore size modulates adipogenic differentiation of MSCs toward identifying optimal scaffold pore size and whether pore size modulates spatial infiltration of adipogenically differentiated cells. To assess this, extrusion‐based 3D printing is used to fabricate photo‐crosslinkable gelatin‐based scaffolds with pore sizes in the range of 200–600 µm. The adipogenic differentiation of MSCs seeded onto these scaffolds is evaluated and robust lipid droplet formation is observed across all scaffold groups as early as after day 6 of culture. Expression of adipogenic genes on scaffolds increases significantly over time, compared to TCP controls. Furthermore, it is found that the spatial distribution of cells is dependent on the scaffold pore size, with larger pores leading to a more uniform spatial distribution of adipogenically differentiated cells. Overall, these data provide first insights into the role of scaffold pore size on MSC‐based adipogenic differentiation and contribute toward the rational design of biomaterials for adipose tissue engineering in 3D volumetric spaces. 相似文献
35.
Klaus Albert Markus Dachtler Tobias Glaser Heidi Hndel Tanja Lacker Gtz Schlotterbeck Sabine Strohschein Li-Hong Tseng Ulrich Braumann 《Journal of separation science》1999,22(3):135-143
The hyphenation of chromatographic separation techniques with NMR spectroscopy is one of the most powerful and time-saving methods for the separation and structural elucidation of unknown compounds and molecular compositions of mixtures. Most of the routinely used NMR flow-cells have detection volumes between 40–180 μL for conventional separations with analytical columns, and the newest designs employ detection volumes in the order of 200 nL for capillary separations. The low flow rates used in capillary chromatography permit the use of deuterated solvents. Unequivocal structural assignment of unknown chromatographic peaks is possible by two-dimensional stopped-flow capillary HPLC-NMR experiments. 相似文献
36.
Shallow Flows Over a Permeable Medium: The Hydrodynamics of Submerged Aquatic Canopies 总被引:2,自引:0,他引:2
Aquatic flow over a submerged vegetation canopy is a ubiquitous example of flow adjacent to a permeable medium. Aquatic canopy
flows, however, have two important distinguishing features. Firstly, submerged vegetation typically grows in shallow regions.
Consequently, the roughness sublayer, the region where the drag length scale of the canopy is dynamically important, can often
encompass the entire flow depth. In such shallow flows, vortices generated by the inflectional velocity profile are the dominant
mixing mechanism. Vertical transport across the canopy–water interface occurs over a narrow frequency range centered around
f
v
(the frequency of vortex passage), with the vortices responsible for more than three-quarters of the interfacial flux. Secondly,
submerged canopies are typically flexible, coupling the motion of the fluid and canopy. Importantly, flexible canopies can
exhibit a coherent waving (the monami) in response to vortex passage. This waving reduces canopy drag, allowing greater in-canopy velocities and turbulent stresses.
As a result, the waving of an experimental canopy reduces the canopy residence time by a factor of four. Finally, the length
required for the set-up and full development of mixing-layer-type canopy flow is investigated. This distance, which scales
upon the drag length scale, can be of the same order as the length of the canopy. In several flows adjacent to permeable media
(such as urban canopies and reef systems), patchiness of the medium is common such that the fully developed condition may
not be representative of the flow as a whole.
This paper has previously been published in Transp Porous Med (2009) 78: 309–326; DOI . 相似文献
37.
Lise-Lotte Gundersen Carl Henrik G?rbitz Liina Neier Heidi Roggen Toomas Tamm 《Theoretical chemistry accounts》2011,129(3-5):349-358
Density Functional Theory calculations of nine 2-substituted N-methoxy-9-methyl-9H-purin-6-amines in the amino and imino tautomeric forms, as well as the complexes of the same with dimethyl sulfoxide (DMSO), were performed using two functionals (BP86 and B3LYP) and two basis sets (SV(P) and def2-TZVP). Solid-state structures of two of the compounds were obtained from single-crystal X-ray diffraction techniques. It was found that the inclusion of both an explicit hydrogen-bonding partner (DMSO) as well as continuum solvation effects, and vibrational corrections to energy, were necessary for qualitative and reasonable quantitative agreement with observed tautomeric ratios. The solution-optimized geometries and X-ray structures were found to be in good agreement. NMR spectroscopy confirmed the dependence of the tautomeric ratios on hydrogen-bonding abilities, in addition to the dipole moment of the solvent in question. Natural Bond Orbital charges on the N-7 nitrogen, as well as the tautomeric ratios were used to explain the observed reactivities of the compounds toward N-7 alkylation. 相似文献
38.
Ane Funderud Heidi H Henanger Tilahun T Hafte Paul S Amieux Sigurd ?rstavik Bj?rn S Sk?lhegg 《BMC biochemistry》2006,7(1):20
Background
Two main genes encoding the catalytic subunits Cα and Cβ of cyclic AMP dependent protein kinase (PKA) have been identified in all vertebrates examined. The murine, bovine and human Cβ genes encode several splice variants, including the splice variant Cβ2. In mouse Cβ2 has a relative molecular mass of 38 kDa and is only expressed in the brain. In human and bovine Cβ2 has a relative molecular mass of 47 kDa and is mainly expressed in lymphoid tissues. 相似文献39.
Dried blood spot (DBS) technology is an emerging alternative for sample collection in bioanalysis. Dilution for DBS samples is a challenge due to its solid sample format. Currently, DBS samples requiring dilution were first extracted as regular samples and then diluted with extracted blank samples containing internal standard (IS). Since the dilution step is a volume-critical step, extra care has to be taken to achieve accurate dilution when dealing with limited volume extracted samples. Here, we introduce an alternative sample dilution for liquid chromatography/tandem mass spectrometry (LC/MS/MS) assays using IS to track the dilution step. Dilution factor-adjusted IS working solution was first added to the sample requiring dilution before sample processing; subsequently, the processed sample was approximately diluted into the assay linear response range before LC/MS/MS analysis. We define this approach as "IS-tracked dilution". The advantage of this approach is that the diluting step is tracked by the IS and is no longer a volume-critical step. Another recognized challenge related to sample dilution is automatic sample dilution using a liquid handler. This "IS-tracked dilution" may also help address some of the challenges for automatic sample dilution of liquid samples. This new dilution approach was proven to be effective and convenient in both plasma assays and DBS assays using omeprazole as a probe compound. 相似文献
40.