Comparing the intracellular fate of components within a noncovalent streptavidin nanoparticle with covalent conjugation
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Authors
Liu, YuxiaCheng, Dengfeng
Liu, Xinrong
Liu, Guozheng
Dou, Shuping
Xiao, Nan
Chen, Ling
Rusckowski, Mary
Hnatowich, Donald J.
Document Type
Journal ArticlePublication Date
2012-01-01Keywords
Antibodies, Monoclonal, HumanizedAntineoplastic Agents
Biotin
Cell Line, Tumor
Drug Delivery Systems
Flow Cytometry
Humans
Indicators and Reagents
Indium Radioisotopes
Microscopy, Fluorescence
Nanoparticles
Neoplasms
Streptavidin
Vitamin B Complex
Nanoparticle
Antisense oligomer
Herceptin
Streptavidin
Cancer Biology
Medicinal-Pharmaceutical Chemistry
Nanomedicine
Oncology
Radiochemistry
Radiology
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Show full item recordAbstract
INTRODUCTION: Auger radiotherapy requires adequate tumor delivery and high nuclear accumulation and retention. We hypothesize that the noncovalent nature of a streptavidin/biotin three-component nanoparticle possessing these qualities may be required for dissociation of the radiolabeled oligomer and its accumulation into the cell nucleus. METHODS: As a test of our hypothesis, the intracellular fate of an antisense oligomer when incubated as the nanoparticle and when incubated while covalently conjugated to the antibody was compared. The three-component noncovalent nanoparticle consisted of streptavidin linking three biotinylated components: a Cy3-labeled anti-RIalpha antisense phosphorodiamidate morpholino (MORF) oligomer, a tat transfecting peptide and the anti-Her2 herceptin antibody. The covalent constructs included an anti-RIalpha antisense DNA conjugated to a radiolabeled herceptin and a fluorescent DNA conjugated to native herceptin. Fluorescence microscopy in SK-BR-3 (Her2+) cells was used to evaluate the fate of the fluorescent Cy5.5-DNA and Cy3-MORF, while the subcellular accumulation of the (111)In-labeled herceptin and herceptin-DNA in both SK-BR-3 and MDA-MB-231 (Her2) cells was determined by isolating and counting the nuclear fractions. RESULTS: Previously, we demonstrated that when incubated as the three-component nanoparticle consisting of herceptin and streptavidin and (99m)Tc-labeled antisense MORF, only the MORF accumulated in the nucleus of Her2+ cells. In this investigation, clear evidence was observed of nuclear accumulation of the antisense oligomer within the noncovalent nanoparticle as before, but when incubated as the covalent construct, by both fluorescence microscopy and nuclear counting, no evidence of nuclear accumulation was observed. CONCLUSION: The weaker noncovalent biotin-streptavidin bond may be essential for adequate delivery of the radiolabeled antisense oligomer to the nucleus of tumor cells.Source
Nucl Med Biol. 2012 Jan;39(1):101-7. Link to article on publisher's siteDOI
10.1016/j.nucmedbio.2011.06.006Permanent Link to this Item
http://hdl.handle.net/20.500.14038/48320PubMed ID
21958854Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1016/j.nucmedbio.2011.06.006