PEGylation is an attractive approach to modifying oligonucleotides intended for therapeutic purposes. PEG conjugation reduces protein interactions with the oligonucleotide, and helps to overcome their intrinsic biopharmaceutical shortcomings, such as poor enzymatic stability, rapid body clearance, and unwanted immunostimulation. However, the effect of PEG architecture and the manner in which the PEG component interferes with the hybridization of the oligonucleotide remain poorly understood. In this study, we systematically compare the hybridization thermodynamics and protein accessibility of several DNA conjugates involving linear, Y-shaped, and brush-type PEG. It is found that PEGylated DNA experiences two opposing effects: local excluded volume effect and chemical interactions, the strengths of which are architecture-dependent. Notably, the brush architecture is able to offer significantly greater protein shielding capacity than its linear or Y-shaped counterparts, while maintaining nearly identical free energy for DNA hybridization compared with free DNA.Opposing effects: The hybridization thermodynamics and protein accessibility of a series of PEG–DNA conjugates were compared. The DNA was found to experience two effects from the PEG component: an excluded volume effect and a chemical interaction effect, which work in opposite directions on duplex stability and are polymer-architecture-dependent.
[NMR paper] Quantitative Detection of Pegylated Biomacromolecules in Biological Fluids by NMR.
Quantitative Detection of Pegylated Biomacromolecules in Biological Fluids by NMR.
Quantitative Detection of Pegylated Biomacromolecules in Biological Fluids by NMR.
Anal Chem. 2016 Feb 29;
Authors: Alvares RD, Hasabnis A, Prosser RS, Macdonald PM
Abstract
The accumulation, bio-distribution and clearance profiles of therapeutic agents are key factors relevant to their efficacy. Determining these properties constitutes an ongoing experimental challenge. Many such therapeutics, including small molecules, peptides, proteins, tissue...
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03-02-2016 07:20 PM
[NMR paper] Solid-State NMR of PEGylated Proteins.
Solid-State NMR of PEGylated Proteins.
Related Articles Solid-State NMR of PEGylated Proteins.
Angew Chem Int Ed Engl. 2016 Jan 12;
Authors: Ravera E, Ciambellotti S, Cerofolini L, Martelli T, Kozyreva T, Bernacchioni C, Giuntini S, Fragai M, Turano P, Luchinat C
Abstract
PEGylated proteins are widely used in biomedicine but, in spite of their importance, no atomic-level information is available since they are generally resistant to structural characterization approaches. PEGylated proteins are shown here to yield highly resolved...
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01-13-2016 05:24 PM
Study of PEGylated model protein reveals porous structure based on PEG size - Phys.Org
Study of PEGylated model protein reveals porous structure based on PEG size - Phys.Org
http://www.bionmr.com//t2.gstatic.com/images?q=tbn:ANd9GcQdT1VtDs_349GghNDjMV50tdw7g0UzZNnkRkMt-SgVH2a6hHSvjT8JY4N42n5PGHBQ_JntWNc
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Study of PEGylated model protein reveals porous structure based on PEG size
Phys.Org
NMR studies using 1H-15N heteronuclear single-quantum correlation spectroscopy showed that the PEG-Pc had well-dispersed resonances that indicated the protein remained folded in a stable conformation. Chemical shift perturbations were only...
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09-22-2015 06:40 PM
A Delicate Interplay of Structure, Dynamics, and Thermodynamics for Function: A High Pressure NMR Study of Outer Surface Protein A
A Delicate Interplay of Structure, Dynamics, and Thermodynamics for Function: A High Pressure NMR Study of Outer Surface Protein A
22 February 2012
Publication year: 2012
Source:Biophysical Journal, Volume 102, Issue 4</br>
</br>
Outer surface protein A (OspA) is a crucial protein in the infection of Borrelia burgdorferi causing Lyme disease. We studied conformational fluctuations of OspA with high-pressure 15N/1H two-dimensional NMR along with high-pressure fluorescence spectroscopy. We found evidence within folded, native OspA for rapid local fluctuations of the...
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02-03-2013 10:13 AM
Erratum to: Paramagnetic labelling of proteins and oligonucleotides for NMR
Erratum to: Paramagnetic labelling of proteins and oligonucleotides for NMR
Erratum to: Paramagnetic labelling of proteins and oligonucleotides for NMR
Content Type Journal Article
Pages 1-2
DOI 10.1007/s10858-011-9475-7
Authors
[NMR paper] NMR studies of protein hydration and TEMPOL accessibility.
NMR studies of protein hydration and TEMPOL accessibility.
Related Articles NMR studies of protein hydration and TEMPOL accessibility.
J Mol Biol. 2003 Sep 12;332(2):437-47
Authors: Niccolai N, Spiga O, Bernini A, Scarselli M, Ciutti A, Fiaschi I, Chiellini S, Molinari H, Temussi PA
Understanding the mechanisms of the interaction between a protein surface and its outer molecular environment is of primary relevance for the rational design of new drugs and engineered proteins. Protein surface accessibility is emerging as a new dimension of...
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11-24-2010 09:16 PM
[NMR paper] NMR studies of protein surface accessibility.
NMR studies of protein surface accessibility.
Related Articles NMR studies of protein surface accessibility.
J Biol Chem. 2001 Nov 9;276(45):42455-61
Authors: Niccolai N, Ciutti A, Spiga O, Scarselli M, Bernini A, Bracci L, Di Maro D, Dalvit C, Molinari H, Esposito G, Temussi PA
Characterization of protein surface accessibility represents a new frontier of structural biology. A surface accessibility investigation for two structurally well-defined proteins, tendamistat and bovine pancreatic trypsin inhibitor, is performed here by a combined...
Effect of PEG Architecture on the Hybridization Thermodynamics and Protein Accessibility of PEGylated Oligonucleotides
Linear Mode
