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Since most recombinant monoclonal antibodies (mAbs) contain glutamic acid or glutamate at their N-terminus, cyclization of these residues to form pyroglutamate is an important degradation pathway that often occurs in therapeutic mAb development. In this work, a rapid method was developed to determine pyroglutamate at the N-terminus of mAb light chain by liquid chromatography coupled with electrospray ionization on a quadrupole time-of-flight mass spectrometer (QTOF). High levels of pyroglutamate were found at the N-terminus of the light chain of a typical recombinant mAb. The quantitative results were comparable to those obtained with a more conventional peptide mapping method. The direct method outlined here can be used to evaluate the impact of N-terminal cyclization during the processing of recombinant mAbs. 相似文献
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The functional Ito formula, in the form df() = f( + d ) f(),is formulated and proved in the context of a Lie algebra L associatedwith a quantum (non-commutative) stochastic calculus. Here fis an element of the universal enveloping algebra U of L, andf() + d() f() is given a meaning using the coproductstructure of U even though the individual terms of this expressionhave no meaning. The Ito formula is equivalent to a chaoticexpansion formula for f() which is found explicitly. 1991 MathematicsSubject Classification: primary 81S25; secondary 60H05; tertiary18B25. 相似文献
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Remmele RL Zhang-van Enk J Dharmavaram V Balaban D Durst M Shoshitaishvili A Rand H 《Journal of the American Chemical Society》2005,127(23):8328-8339
The role of thermal unfolding as it pertains to thermodynamic properties of proteins and their stability has been the subject of study for more than 50 years. Moreover, exactly how the unfolding properties of a given protein system may influence the kinetics of aggregation has not been fully characterized. In the study of recombinant human Interleukin-1 receptor type II (rhuIL-1R(II)) aggregation, data obtained from size exclusion chromatography and differential scanning calorimetry (DSC) were used to model the thermodynamic and kinetic properties of irreversible denaturation. A break from linearity in the initial aggregation rates as a function of 1/T was observed in the vicinity of the melting transition temperature (T(m) approximately 53.5 degrees C), suggesting significant involvement of protein unfolding in the reaction pathway. A scan-rate dependence in the DSC experiment testifies to the nonequilibrium influences of the aggregation process. A mechanistic model was developed to extract meaningful thermodynamic and kinetic parameters from an irreversibly denatured process. The model was used to simulate how unfolding properties could be used to predict aggregation rates at different temperatures above and below the T(m) and to account for concentration dependence of reaction rates. The model was shown to uniquely identify the thermodynamic parameters DeltaC(P) (1.3 +/- 0.7 kcal/mol-K), DeltaH(m) (74.3 +/- 6.8 kcal/mol), and T(m) with reasonable variances. 相似文献
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Lymperakis L Neugebauer J Albrecht M Remmele T Strunk HP 《Physical review letters》2004,93(19):196401
Combining through-focus high-resolution transmission electron microscopy and hierarchical multiscale simulations consisting of density-functional theory, analytical empirical potentials, and continuum elastic theory we demonstrate the existence of a new dislocation type in GaN. In contrast with all previously identified or suggested dislocation structures in GaN, all core atoms are fully coordinated; i.e., no broken bonds occur, implying that the dislocation should be electrically inactive. However, as we show, the giant local strain-field around the dislocation core, in combination with the small lattice constant of GaN, causes deep defect states and thus electrically active edge dislocations independent on the specific core structure. 相似文献
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Bobeldijk I Bouwhuis M Ireland DG de Jager CW Jans E de Jonge N Kasdorp WJ Konijn J Lapikás L van Leeuwe JJ van der Meer RL Nooren GJ Passchier E Schroevers M van der Steenhoven G Steijger JJ Theunissen JA van Uden MA de Vries H de Vries R de Witt Huberts PK Blok HP van den Brink HB Dodge GE Harakeh MN Hesselink WH Kalantar-Nayestanaki N Pellegrino A Spaltro CM Templon JA Hicks RS Kelly JJ Marchand C 《Physical review letters》1994,73(20):2684-2687
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