Limits...
PET/CT Based In Vivo Evaluation of 64Cu Labelled Nanodiscs in Tumor Bearing Mice.

Huda P, Binderup T, Pedersen MC, Midtgaard SR, Elema DR, Kjær A, Jensen M, Arleth L - PLoS ONE (2015)

Bottom Line: This makes them promising candidates for both drug delivery purposes and as advanced imaging agents.For the radiolabelling, a simple approach for 64Cu radiolabelling of proteins via a chelating agent, DOTA, was developed.The reaction was performed at sufficiently mild conditions to be compatible with labelling of the protein part of a lipid-protein particle while fully conserving the particle structure including the amphipathic protein fold.

View Article: PubMed Central - PubMed

Affiliation: Structural Biophysics, Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, DK-2100 København Ø, Denmark.

ABSTRACT
64Cu radiolabelled nanodiscs based on the 11 α-helix MSP1E3D1 protein and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine lipids were, for the first time, followed in vivo by positron emission tomography for evaluating the biodistribution of nanodiscs. A cancer tumor bearing mouse model was used for the investigations, and it was found that the approximately 13 nm nanodiscs, due to their size, permeate deeply into cancer tissue. This makes them promising candidates for both drug delivery purposes and as advanced imaging agents. For the radiolabelling, a simple approach for 64Cu radiolabelling of proteins via a chelating agent, DOTA, was developed. The reaction was performed at sufficiently mild conditions to be compatible with labelling of the protein part of a lipid-protein particle while fully conserving the particle structure including the amphipathic protein fold.

No MeSH data available.


Related in: MedlinePlus

Purification of 64Cu-nanodiscs.Chromatogram obtained by size exclusion chromatography using a Superdex 200 (10/300) column detected by UV 280 nm absorption (mAU) and radioactivity at 511 kEV (intensity). 64Cu-nanodiscs were collected from the column at an elution time of 12.4 ml.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4488450&req=5

pone.0129310.g003: Purification of 64Cu-nanodiscs.Chromatogram obtained by size exclusion chromatography using a Superdex 200 (10/300) column detected by UV 280 nm absorption (mAU) and radioactivity at 511 kEV (intensity). 64Cu-nanodiscs were collected from the column at an elution time of 12.4 ml.

Mentions: DOTA-modified nanodiscs were 64Cu-labelled by simple incubation with 64CuCl2 in a pH 7.5 ammonium acetate buffer at room temperature. The labelling was confirmed by SEC with dual detection of UV 280 nm and gamma-emission, which clearly showed that 64Cu was bound to the nanodisc fraction (Fig 3). The finding of efficient radiolabelling at these mild conditions was very satisfactory, as previous work reports the requirement for more harsh reaction conditions at lower pH [23–26], which is generally not compatible with the conservation of the structure and function of protein-based biomolecules.


PET/CT Based In Vivo Evaluation of 64Cu Labelled Nanodiscs in Tumor Bearing Mice.

Huda P, Binderup T, Pedersen MC, Midtgaard SR, Elema DR, Kjær A, Jensen M, Arleth L - PLoS ONE (2015)

Purification of 64Cu-nanodiscs.Chromatogram obtained by size exclusion chromatography using a Superdex 200 (10/300) column detected by UV 280 nm absorption (mAU) and radioactivity at 511 kEV (intensity). 64Cu-nanodiscs were collected from the column at an elution time of 12.4 ml.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4488450&req=5

pone.0129310.g003: Purification of 64Cu-nanodiscs.Chromatogram obtained by size exclusion chromatography using a Superdex 200 (10/300) column detected by UV 280 nm absorption (mAU) and radioactivity at 511 kEV (intensity). 64Cu-nanodiscs were collected from the column at an elution time of 12.4 ml.
Mentions: DOTA-modified nanodiscs were 64Cu-labelled by simple incubation with 64CuCl2 in a pH 7.5 ammonium acetate buffer at room temperature. The labelling was confirmed by SEC with dual detection of UV 280 nm and gamma-emission, which clearly showed that 64Cu was bound to the nanodisc fraction (Fig 3). The finding of efficient radiolabelling at these mild conditions was very satisfactory, as previous work reports the requirement for more harsh reaction conditions at lower pH [23–26], which is generally not compatible with the conservation of the structure and function of protein-based biomolecules.

Bottom Line: This makes them promising candidates for both drug delivery purposes and as advanced imaging agents.For the radiolabelling, a simple approach for 64Cu radiolabelling of proteins via a chelating agent, DOTA, was developed.The reaction was performed at sufficiently mild conditions to be compatible with labelling of the protein part of a lipid-protein particle while fully conserving the particle structure including the amphipathic protein fold.

View Article: PubMed Central - PubMed

Affiliation: Structural Biophysics, Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, DK-2100 København Ø, Denmark.

ABSTRACT
64Cu radiolabelled nanodiscs based on the 11 α-helix MSP1E3D1 protein and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine lipids were, for the first time, followed in vivo by positron emission tomography for evaluating the biodistribution of nanodiscs. A cancer tumor bearing mouse model was used for the investigations, and it was found that the approximately 13 nm nanodiscs, due to their size, permeate deeply into cancer tissue. This makes them promising candidates for both drug delivery purposes and as advanced imaging agents. For the radiolabelling, a simple approach for 64Cu radiolabelling of proteins via a chelating agent, DOTA, was developed. The reaction was performed at sufficiently mild conditions to be compatible with labelling of the protein part of a lipid-protein particle while fully conserving the particle structure including the amphipathic protein fold.

No MeSH data available.


Related in: MedlinePlus