[NMR paper] Strategies for the production of isotopically labelled Fab fragments of therapeutic antibodies in Komagataella phaffii (Pichia pastoris) and Escherichia coli for NMR studies
Strategies for the production of isotopically labelled Fab fragments of therapeutic antibodies in Komagataella phaffii (Pichia pastoris) and Escherichia coli for NMR studies
The importance and fast growth of therapeutic monoclonal antibodies, both innovator and biosimilar products, have triggered the need for the development of characterization methods at high resolution such as nuclear magnetic resonance (NMR) spectroscopy. However, the full power of NMR spectroscopy cannot be unleashed without labelling the mAb of interest with NMR-active isotopes. Here, we present strategies using either Komagataella phaffii (Pichia pastoris) or Escherichia coli that can be...
[NMR paper] Production and Preparation of Isotopically Labeled Human Membrane Proteins in Pichia pastoris for Fast-MAS-NMR Analyses
Production and Preparation of Isotopically Labeled Human Membrane Proteins in Pichia pastoris for Fast-MAS-NMR Analyses
Membrane proteins (MPs) comprise about one-third of the human proteome, playing critical roles in many physiological processes and associated disorders. Consistently, they represent one of the largest classes of targets for the pharmaceutical industry. Their study at the molecular level is however particularly challenging, resulting in a severe lack of structural and dynamic information that is hindering their detailed functional characterization and the identification of...
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07-01-2022 10:59 AM
[NMR paper] Improved Protocol for the Production of the Low-Expression Eukaryotic Membrane Protein Human Aquaporin 2 in Pichia pastoris for Solid-State NMR.
Improved Protocol for the Production of the Low-Expression Eukaryotic Membrane Protein Human Aquaporin 2 in Pichia pastoris for Solid-State NMR.
Related Articles Improved Protocol for the Production of the Low-Expression Eukaryotic Membrane Protein Human Aquaporin 2 in Pichia pastoris for Solid-State NMR.
Biomolecules. 2020 Mar 11;10(3):
Authors: Munro R, de Vlugt J, Ladizhansky V, Brown LS
Abstract
Solid-state nuclear magnetic resonance (SSNMR) is a powerful biophysical technique for studies of membrane proteins; it requires the...
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03-16-2020 04:59 PM
Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
Overproduction of 15N-labeled r-RGD-hirudin in pichia pastoris for NMR studies.
Protein Pept Lett. 2010;17(10):1228-33
Authors: Wang J, Zhang Y, Li S, Liu X, Yan X, Song H, Yu M, Dai L, Mo W
The novel recombinant hirudin, r-RGD-hirudin, inhibits thrombin and platelet aggregation. Here, we reported over-expression of (15)N-labeled r-RGD-hirudin by Pichia pastoris in minimal medium. After extensive optimization, the yield of active r-RGD-hirudin reached ?600 mg/L when...
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Proteins
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01-22-2011 01:52 PM
Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells
Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells
Abstract The preparation of stable isotope-labeled proteins is necessary for the application of a wide variety of NMR methods, to study the structures and dynamics of proteins and protein complexes. The E. coli expression system is generally used for the production of isotope-labeled proteins, because of the advantages of ease of handling, rapid growth, high-level protein production, and low cost for isotope-labeling. However, many eukaryotic proteins are not functionally expressed...
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01-09-2011 12:46 PM
[NMR paper] Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR
Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR studies.
Related Articles Overexpression and purification of isotopically labeled Escherichia coli MutH for NMR studies.
Protein Expr Purif. 2003 Jun;29(2):252-8
Authors: Dutta A, Rao BJ, Chary KV
MutH is one of the enzymes involved in the methyl directed -GATC-based DNA repair system. We report a significantly optimized protocol to prepare isotopically (15N and/or 13C) labeled MutH in minimal medium with high yields for NMR studies. Under the various...
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11-24-2010 09:01 PM
[NMR paper] Expression of deuterium-isotope-labelled protein in the yeast pichia pastoris for NMR
Expression of deuterium-isotope-labelled protein in the yeast pichia pastoris for NMR studies.
Related Articles Expression of deuterium-isotope-labelled protein in the yeast pichia pastoris for NMR studies.
J Biomol NMR. 2000 Aug;17(4):337-47
Authors: Morgan WD, Kragt A, Feeney J
Deuterium isotope labelling is important for NMR studies of large proteins and complexes. Many eukaryotic proteins are difficult to express in bacteria, but can be efficiently produced in the methylotrophic yeast Pichia pastoris. In order to facilitate NMR studies of...
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11-19-2010 08:29 PM
[NMR paper] Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pa
Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris.
Related Articles Isotopically labeled bovine beta-lactoglobulin for NMR studies expressed in Pichia pastoris.
Protein Expr Purif. 1998 Oct;14(1):97-103
Authors: Denton H, Smith M, Husi H, Uhrin D, Barlow PN, Batt CA, Sawyer L
beta-Lactoglobulin (beta-Lg) is the major whey protein in ruminant milk and has been implicated in the irreversible denaturation of milk proteins and its associated poor processing behavior during heat treatment. In order to help...
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11-17-2010 11:15 PM
Production of recombinant isotopically labelled peptide by fusion to an insoluble par
Production of recombinant isotopically labelled peptide by fusion to an insoluble partner protein: generation of integrin ?v?6 binding peptides for NMR.
Related Articles Production of recombinant isotopically labelled peptide by fusion to an insoluble partner protein: generation of integrin ?v?6 binding peptides for NMR.
Mol Biosyst. 2010 Oct 18;
Authors: Wagstaff JL, Howard MJ, Williamson RA
The integrin ?v?6 is up-regulated in several cancers and has clinical potential for both tumour imaging and therapy. Peptide ligands have been developed...
Strategies for the production of isotopically labelled Fab fragments of therapeutic antibodies in Komagataella phaffii (Pichia pastoris) and Escherichia coli for NMR studies
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