Related ArticlesProbing Microenvironmental Acidity in Lyophilized Protein and Vaccine Formulations Using Solid-state NMR Spectroscopy.
J Pharm Sci. 2020 Nov 26;:
Authors: Li M, Koranne S, Fang R, Lu X, Williams DM, Munson EJ, Bhambhani A, Su Y
Abstract
Biophysical and biochemical instability of therapeutic proteins in the solution state may necessitate the development of products in the solid form, due to their enhanced stability. Lyophilization is a widely used method to ensure dry state stabilization of biological products. A commonly encountered issue is the pH shifts that can occur due to undesired crystallization of a buffer component, resulting in loss of protein activities. However, it is technically challenging to noninvasively investigate the physicochemical environment in the lyophile matrix. In this work, we demonstrate an approach based on solid-state NMR to investigate the microenvironmental acidity in lyophilized protein formulations, using histidine, a commonly used buffer agent, as a molecular probe. The solid-state acidity in the lyophilized matrix can be assessed by monitoring the chemical shift changes of histidine. The protonation and tautomeric states of histidine lyophilized at a range of pH values from 4.5 to 11.0 were identified from full 13C and 15N resonance assignments in one-dimensional and two-dimensional NMR experiments. The results demonstrated a pH-dependence of histidine chemical shift in the amorphous state. Moreover, we successfully applied this protocol to investigate the microenvironmental pH in lyophilized formulations of the HPV vaccine and lactate dehydrogenase protein.
PMID: 33249049 [PubMed - as supplied by publisher]
Probing Functionalities and Acidity of Calcined Phenylene-Bridged Periodic Mesoporous Organosilicates Using (DNP)-NMR, DRIFTS and XPS #DNPNMR
From The DNP-NMR Blog:
Probing Functionalities and Acidity of Calcined Phenylene-Bridged Periodic Mesoporous Organosilicates Using (DNP)-NMR, DRIFTS and XPS #DNPNMR
Pirez, Cyril, Hiroki Nagashima, Franck Dumeignil, and Olivier Lafon. “Probing Functionalities and Acidity of Calcined Phenylene-Bridged Periodic Mesoporous Organosilicates Using (DNP)-NMR, DRIFTS and XPS.” The Journal of Physical Chemistry C, February 19, 2020, acs.jpcc.9b11223.
https://doi.org/10.1021/acs.jpcc.9b11223.
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03-16-2020 04:59 PM
[NMR paper] Acidity Strength of Solid Catalysts Probed by Hyperpolarized Natural Abundance 17O NMR Spectroscopy
Acidity Strength of Solid Catalysts Probed by Hyperpolarized Natural Abundance 17O NMR Spectroscopy
Observation of Brønsted acid sites: By enhancing the solid-state NMR signals of 17O at natural abundance with dynamic nuclear polarization (DNP), Pruski et al. were able to measure oxygen–proton distances accurately with sub-picometer precision. The results give insight into the Brønsted acidity of a range of solid acid catalysts.
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07-11-2017 09:20 AM
[NMR paper] Probing the Conformationally Excited States of Membrane Proteins via (1)H-detected MAS Solid-State NMR Spectroscopy.
Probing the Conformationally Excited States of Membrane Proteins via (1)H-detected MAS Solid-State NMR Spectroscopy.
Probing the Conformationally Excited States of Membrane Proteins via (1)H-detected MAS Solid-State NMR Spectroscopy.
J Phys Chem B. 2017 Apr 13;:
Authors: Gopinath T, Nelson SE, Soller KJ, Veglia G
Abstract
Proteins exist in ensembles of conformational states that interconvert on various motional time scales. High-energy states of proteins, often referred to as conformationally excited states, are sparsely...
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04-14-2017 10:27 AM
[NMR paper] Probing Transient Conformational States of Proteins by Solid-State R1? Relaxation-Dispersion NMR Spectroscopy.
Probing Transient Conformational States of Proteins by Solid-State R1? Relaxation-Dispersion NMR Spectroscopy.
Related Articles Probing Transient Conformational States of Proteins by Solid-State R1? Relaxation-Dispersion NMR Spectroscopy.
Angew Chem Int Ed Engl. 2014 Mar 18;
Authors: Ma P, Haller JD, Zajakala J, Macek P, Sivertsen AC, Willbold D, Boisbouvier J, Schanda P
Abstract
The function of proteins depends on their ability to sample a variety of states differing in structure and free energy. Deciphering how the various thermally...
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03-20-2014 12:44 PM
[NMR paper] Probing the Interaction of Arg9Cys Mutated Phospholamban with Phospholipid Bilayers by Solid-state NMR Spectroscopy.
Probing the Interaction of Arg9Cys Mutated Phospholamban with Phospholipid Bilayers by Solid-state NMR Spectroscopy.
Related Articles Probing the Interaction of Arg9Cys Mutated Phospholamban with Phospholipid Bilayers by Solid-state NMR Spectroscopy.
Biochim Biophys Acta. 2013 Jul 9;
Authors: Yu X, Lorigan GA
Abstract
Phospholamban (PLB) is a 52 amino acid integral membrane protein that interacts with the sarcoplasmic reticulum Ca(2+) ATPase (SERCA) and helps to regulate Ca(2+) flow. PLB inhibits SERCA impairing Ca(2+) translocation. The...
[NMR paper] A solid-state NMR study of protein mobility in lyophilized protein-sugar powders.
A solid-state NMR study of protein mobility in lyophilized protein-sugar powders.
Related Articles A solid-state NMR study of protein mobility in lyophilized protein-sugar powders.
J Pharm Sci. 2002 Apr;91(4):943-51
Authors: Lam YH, Bustami R, Phan T, Chan HK, Separovic F
The molecular mobility of protein in lyophilized lysozyme-sugar systems stored at different relative humidities was studied using solid-state NMR. Relaxation measurements, T(1) of high-frequency (MHz), and T(1rho), of low-frequency (kHz) motions, were performed on lysozyme...
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11-24-2010 08:49 PM
[NMR paper] Molecular mobility of protein in lyophilized formulations linked to the molecular mob
Molecular mobility of protein in lyophilized formulations linked to the molecular mobility of polymer excipients, as determined by high resolution 13C solid-state NMR.
Related Articles Molecular mobility of protein in lyophilized formulations linked to the molecular mobility of polymer excipients, as determined by high resolution 13C solid-state NMR.
Pharm Res. 1999 Oct;16(10):1621-5
Authors: Yoshioka S, Aso Y, Kojima S, Sakurai S, Fujiwara T, Akutsu H
PURPOSE: The mobility of protein molecules in lyophilized protein formulations was compared...
Probing Microenvironmental Acidity in Lyophilized Protein and Vaccine Formulations Using Solid-state NMR Spectroscopy.
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