Labeling of adenovirus particles with PARACEST agents.
Identifieur interne : 000597 ( PubMed/Corpus ); précédent : 000596; suivant : 000598Labeling of adenovirus particles with PARACEST agents.
Auteurs : Olga Vasalatiy ; Robert D. Gerard ; Piyu Zhao ; Xiankai Sun ; A Dean SherrySource :
- Bioconjugate chemistry [ 1043-1802 ] ; 2008.
English descriptors
- KwdEn :
- Adenoviridae (chemistry), Adenoviridae (genetics), Adenoviridae Infections (genetics), Adenoviridae Infections (virology), Cell Line, Chelating Agents (chemistry), Contrast Media (chemistry), Crown Compounds (chemistry), Humans, Indicators and Reagents, Kinetics, Ligands, Luciferases (genetics), Luminescence, Lutetium, Magnetic Resonance Spectroscopy, Metals (chemistry), Radioisotopes, Spectrometry, Mass, Electrospray Ionization, Thulium.
- MESH :
- chemical , chemistry : Chelating Agents, Contrast Media, Crown Compounds, Metals.
- chemistry : Adenoviridae.
- genetics : Adenoviridae, Adenoviridae Infections, Luciferases.
- virology : Adenoviridae Infections.
- Cell Line, Humans, Indicators and Reagents, Kinetics, Ligands, Luminescence, Lutetium, Magnetic Resonance Spectroscopy, Radioisotopes, Spectrometry, Mass, Electrospray Ionization, Thulium.
Abstract
Recombinant adenovirus type 5 particles (AdCMVLuc) were labeled with two different bifunctional ligands capable of forming stable complexes with paramagnetic lanthanide ions. The number of covalently attached ligands varied between 630 and 1960 per adenovirus particle depending upon the chemical reactivity of the bifunctional ligand (NHS ester versus isothiocyanide), the amount of excess ligand added, and the reaction time. The bioactivity of each labeled adenovirus derivative, as measured by the ability of the virus to infect cells and express luciferase, was shown to be highly dependent upon the number of covalently attached ligands. This indicates that certain amino groups, likely on the surface of the adenovirus fiber protein where cell binding is known to occur, are critical for viral attachment and infection. Addition of (177)Lu3+ to chemically modified versus control viruses demonstrated a significant amount of nonspecific binding of (177)Lu3+ to the virus particles that could not be sequestered by addition of excess DTPA. Thus, it became necessary to implement a prelabeling strategy for conjugation of preformed lanthanide ligand chelates to adenovirus particles. Using preformed Tm3+- L2, a large number of chelates having chemical exchange saturation transfer (CEST) properties were attached to the surface residues of AdCMVLuc without nonspecific binding of metal ions elsewhere on the virus particle. The potential of such conjugates to act as PARACEST imaging agents was tested using an on-resonance WALTZ sequence for CEST activation. A 12% decrease in bulk water signal intensity was observed relative to controls. This demonstrates that viral particles labeled with PARACEST-type imaging agents can potentially serve as targeted agents for molecular imaging.
DOI: 10.1021/bc7002605
PubMed: 18254605
Links to Exploration step
pubmed:18254605Le document en format XML
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<author><name sortKey="Gerard, Robert D" sort="Gerard, Robert D" uniqKey="Gerard R" first="Robert D" last="Gerard">Robert D. Gerard</name>
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<author><name sortKey="Sun, Xiankai" sort="Sun, Xiankai" uniqKey="Sun X" first="Xiankai" last="Sun">Xiankai Sun</name>
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<term>Luciferases (genetics)</term>
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<term>Lutetium</term>
<term>Magnetic Resonance Spectroscopy</term>
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<term>Radioisotopes</term>
<term>Spectrometry, Mass, Electrospray Ionization</term>
<term>Thulium</term>
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<term>Contrast Media</term>
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<term>Indicators and Reagents</term>
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<term>Ligands</term>
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<term>Lutetium</term>
<term>Magnetic Resonance Spectroscopy</term>
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<front><div type="abstract" xml:lang="en">Recombinant adenovirus type 5 particles (AdCMVLuc) were labeled with two different bifunctional ligands capable of forming stable complexes with paramagnetic lanthanide ions. The number of covalently attached ligands varied between 630 and 1960 per adenovirus particle depending upon the chemical reactivity of the bifunctional ligand (NHS ester versus isothiocyanide), the amount of excess ligand added, and the reaction time. The bioactivity of each labeled adenovirus derivative, as measured by the ability of the virus to infect cells and express luciferase, was shown to be highly dependent upon the number of covalently attached ligands. This indicates that certain amino groups, likely on the surface of the adenovirus fiber protein where cell binding is known to occur, are critical for viral attachment and infection. Addition of (177)Lu3+ to chemically modified versus control viruses demonstrated a significant amount of nonspecific binding of (177)Lu3+ to the virus particles that could not be sequestered by addition of excess DTPA. Thus, it became necessary to implement a prelabeling strategy for conjugation of preformed lanthanide ligand chelates to adenovirus particles. Using preformed Tm3+- L2, a large number of chelates having chemical exchange saturation transfer (CEST) properties were attached to the surface residues of AdCMVLuc without nonspecific binding of metal ions elsewhere on the virus particle. The potential of such conjugates to act as PARACEST imaging agents was tested using an on-resonance WALTZ sequence for CEST activation. A 12% decrease in bulk water signal intensity was observed relative to controls. This demonstrates that viral particles labeled with PARACEST-type imaging agents can potentially serve as targeted agents for molecular imaging.</div>
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<Abstract><AbstractText>Recombinant adenovirus type 5 particles (AdCMVLuc) were labeled with two different bifunctional ligands capable of forming stable complexes with paramagnetic lanthanide ions. The number of covalently attached ligands varied between 630 and 1960 per adenovirus particle depending upon the chemical reactivity of the bifunctional ligand (NHS ester versus isothiocyanide), the amount of excess ligand added, and the reaction time. The bioactivity of each labeled adenovirus derivative, as measured by the ability of the virus to infect cells and express luciferase, was shown to be highly dependent upon the number of covalently attached ligands. This indicates that certain amino groups, likely on the surface of the adenovirus fiber protein where cell binding is known to occur, are critical for viral attachment and infection. Addition of (177)Lu3+ to chemically modified versus control viruses demonstrated a significant amount of nonspecific binding of (177)Lu3+ to the virus particles that could not be sequestered by addition of excess DTPA. Thus, it became necessary to implement a prelabeling strategy for conjugation of preformed lanthanide ligand chelates to adenovirus particles. Using preformed Tm3+- L2, a large number of chelates having chemical exchange saturation transfer (CEST) properties were attached to the surface residues of AdCMVLuc without nonspecific binding of metal ions elsewhere on the virus particle. The potential of such conjugates to act as PARACEST imaging agents was tested using an on-resonance WALTZ sequence for CEST activation. A 12% decrease in bulk water signal intensity was observed relative to controls. This demonstrates that viral particles labeled with PARACEST-type imaging agents can potentially serve as targeted agents for molecular imaging.</AbstractText>
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