Water soluble polymers and their derivatives bound to proteins can dramatically favor the biological activity of new drugs and vaccines. Quantification of the modification degree of the protein is crucial during the development and licensing phase and later in order to monitor the industrial production process and to match product specification. In this work, we describe an innovative way to measure directly the modification degree of polysialylated proteins using proton NMR (Nuclear Magnetic...
The NMR signature of gluconoylation: a frequent N-terminal modification of isotope-labeled proteins
The NMR signature of gluconoylation: a frequent N-terminal modification of isotope-labeled proteins
Abstract
N-terminal gluconoylation is a moderately widespread modification in recombinant proteins expressed in Escherichia coli, in particular in proteins bearing an N-terminal histidine-tag. This post-translational modification has been investigated mainly by mass spectrometry. Although its NMR signals must have been observed earlier in spectra of 13C/15N labeled proteins, their chemical shifts were not yet reported. Here we present the complete 1H...
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03-24-2019 10:41 PM
[NMR paper] Palladium in the Chemical Synthesis and Modification of Proteins
Palladium in the Chemical Synthesis and Modification of Proteins
The field of site-specific modification of proteins has drawn significant attention in recent years owing to its importance in various research areas such as the development of novel therapeutics and understanding the biochemical and cellular behaviors of proteins. The presence of a large number of reactive functional groups in the protein of interest and in the cellular environment renders modification at a specific site a highly challenging task. With the development of sophisticated chemical methodologies it is now...
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08-07-2017 07:31 PM
[NMR paper] Heteronuclear NMR As a 4-in-1 Analytical Platform for Detecting Modification-Specific Signatures of Therapeutic Insulin Formulations.
Heteronuclear NMR As a 4-in-1 Analytical Platform for Detecting Modification-Specific Signatures of Therapeutic Insulin Formulations.
Related Articles Heteronuclear NMR As a 4-in-1 Analytical Platform for Detecting Modification-Specific Signatures of Therapeutic Insulin Formulations.
Anal Chem. 2014 Feb 5;
Authors: Jin X, Kang S, Kwon H, Park S
Abstract
Detecting possible modifications of therapeutic proteins is a critical element of the quality control of protein drugs. Typically, a number of techniques are used to evaluate different...
Journal Highlight: Assessment of higher order structure comparability in therapeutic proteins using nuclear magnetic resonance spectroscopy
Journal Highlight: Assessment of higher order structure comparability in therapeutic proteins using nuclear magnetic resonance spectroscopy
http://www.spectroscopynow.com/common/images/thumbnails/13ef9b3d882.jpgNMR spectroscopy using a fingerprinting approach has been used to rapidly assess higher order structure comparability in three nonglycosylated proteins spanning a molecular weight range of 6.5–67 kDa.
Read the rest at Spectroscopynow.com
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06-03-2013 04:21 PM
Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency
Optimal degree of protonation for 1H detection of aliphatic sites in randomly deuterated proteins as a function of the MAS frequency
Abstract The 1H dipolar network, which is the major obstacle for applying proton detection in the solid-state, can be reduced by deuteration, employing the RAP (Reduced Adjoining Protonation) labeling scheme, which yields random protonation at non-exchangeable sites. We present here a systematic study on the optimal degree of random sidechain protonation in RAP samples as a function of the MAS (magic angle spinning) frequency. In particular, we compare 1H...
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08-25-2012 08:56 AM
[U. of Ottawa NMR Facility Blog] Fast 90 Degree Pulse Determination
Fast 90 Degree Pulse Determination
Almost all NMR measurements rely on the correct calibration of 90° pulses. This is traditionally done by collecting a series of spectra as a function of pulse duration, finding a null for the 180° or 360° pulse and calculating the 90° pulse by simple division by 2 or 4 in the case of the 180° and 360° nulls, respectively. This determination, although trivial, can be very time consuming. Wu and Otting* have presented a much faster method of determining a 90° pulse based on measuring the nutation of a magnetization vector directly. Continuous nutation is...
Determination of modification degree of polysialylated therapeutic proteins using (1)H-NMR spectroscopy
Linear Mode
