期刊
BIOCONJUGATE CHEMISTRY
卷 13, 期 1, 页码 122-127出版社
AMER CHEMICAL SOC
DOI: 10.1021/bc0155521
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- NCI NIH HHS [5R01 CA74424-01, 1P50CA86355-01] Funding Source: Medline
- NATIONAL CANCER INSTITUTE [P50CA086355, R01CA074424] Funding Source: NIH RePORTER
Covalent conjugates of the cross-linked iron oxide nanoparticles (CLIO) and high-affinity (K-d(app) = 8.5 nM) anti-human E-selectin (CD62E) F(ab')(2) fragments were prepared and tested in vitro to establish feasibility of endothelial proinflammatory marker magnetic resonance (MR) imaging. The conjugates were obtained by using thiol-disulfide exchange reaction between 3-(2-pyridyl)propionyl-CLIO and S-acetylthioacetate-modified F(ab')(2) fragments. The purified CLIO-F(ab')(2) conjugates (average hydrodynamic diameter 40.6 nm) were used in experiments with the live human endothelial umbilical vein cells (HUVEC). Cells treated with IL-1beta expressed E-selectin and showed a 100-200 times higher binding of CLIO particles (83-104 ng iron/million cells) than control cells. The binding resulted in a high superparamagnetism of HUVEC with the transverse water proton relaxation time (T2) decrease to 30-40 ins in cell precipitates. Cells did not bind/internalize CLIO-F(ab')(2) conjugates prepared using a control fragment or nonconjugated iron oxide particles before or after treatment with IL-1beta. MR imaging of cells showed a highly specific T2-weighted signal darkening associated with cells treated with IL-1beta and incubated with anti-E selectin. Demonstration of MR imaging of E-selectin expression justifies further development of MR-targeted agents for monitoring tumor vascular endothelial proliferation, angiogenesis, and atherosclerosis.
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