)”
“N-glycan analysis of recombinant monoclonal antibodies (mAbs) usually requires the removal of oligosaccharides by PNGase F followed by 2-AB labeling, normal phase high performance liquid chromatography (NP-HPLC) separation and fluorescence detection. Alternatively antibodies can be completely digested by trypsin to generate glycopeptides for analysis by liquid chromatography-mass spectrometry (LC-MS). Here, we report the development of a rapid digestion procedure to generate glycopeptides for quantitative LC-MS analysis. Recombinant monoclonal antibodies were digested using a combination of Lys-C and trypsin at 37 degrees C for 15 min. The glycan profiles from this rapid digestion procedure are in good agreement

with those from LC-MS analysis of glycopeptides from completely digested antibodies and those from NP-HPLC analysis of 2-AB labeled PNGase F released oligosaccharides. This rapid digestion procedure was applied to the comparison of oligosaccharides Fer-1 of two different antibodies. Glycopeptides from the two antibodies were differentially labeled with stable isotopes and analyzed simultaneously after a 1:1 mixing. The combination of the rapid digestion procedure and differential stable isotope labeling significantly www.selleckchem.com/products/i-bet151-gsk1210151a.html reduced the turnaround time. (C) 2012 Elsevier B.V. All rights reserved.”
“Despite

the efforts that bioengineers have exerted in designing and constructing biological processes that function according to a predetermined set of rules, their operation remains fundamentally circumstantial. The contextual situation in which

molecules and single-celled or multi-cellular organisms find p38 MAPK pathway themselves shapes the way they interact, respond to the environment and process external information. Since the birth of the field, synthetic biologists have had to grapple with contextual issues, particularly when the molecular and genetic devices inexplicably fail to function as designed when tested in vivo. In this review, we set out to identify and classify the sources of the unexpected divergences between design and actual function of synthetic systems and analyze possible methodologies aimed at controlling, if not preventing, unwanted contextual issues.”
“Background: The measurement of resting energy expenditure (REE) in an ambulatory setting raises methodological problems. Therefore, the use of predictive equations for the estimation of REE is common. Alternatively, the measurement of sleeping energy expenditure (SEE) has been proposed. The authors retrospectively analyzed data on SEE assessed with a portable armband (PA) device in an ambulatory setting and evaluated this approach against predictive equations and REE measured by indirect calorimetry (IC). Methods: REE was measured with IC, and SEE was assessed with the PA using standardized conditions in 81 participants (aged 46 +/- 13 years) over a wide range of body weight (mean body mass index [BMI] 36.4 +/- 9.3 kg/m(2); range, 21.