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Delivery of DNA Through the CD5 Receptor Using an Avidinbiotin T101 Antibody Conjugate System

Published

Author(s)

M C. Chakrabarti, J A. Carrasquillo, D Drumm, R D. Neumann, I Panyutin

Abstract

We investigated the selective transfer of a plasmid to the target cells using an internalizing monoclonal antibody. We used T101 (IgG2a), a murinemonoclonal antibody that recognizes CD5 antigen, and a plasmid DNA, which contained luciferase reporter gene. Avidin-polylysine was prepared by covalent attachment of polylysine with a carbohydrate chain of avidin via periodate oxidation and sodium borohydride reduction. HPLC purified avidin-polylysine conjugate was reacted with biotinylated T101 and further conjugated with 125I-labeled pGL2 plasmid DNA. The binding between-pGL2 DNA with purified polylysine-avidin-biotin-T101 was studied by nitrocellulose binding and agarose gel retardation assays. We also evaluated the immunoreactivity and retention assay of 125I in the CD5 positive human cells with 125I-pGL2-polylysine-avidin-biotin-T101 conjugates. The 125I-pGL2-polylysine-avidin-biotin-T101 conjugate was 73% immunoreactive and had a prolonged retention of activity by CCRF-CEM cell line at 37 C, whereas 125I-T101 was rapidly released. The intracellular localization studies showed that 80% of radioactivity was associated with the nuclear fraction at 24 h with 125I-pGL2 bound polylysine-avidin-biotin-T101. An in vitro transfection study of CCRF-CEM cell showed that there was a 35-fold greater expression of luciferase gene in the case of pGL2-polylysine-avidin-biotin-T101 than in the control pGL2-polylysine-avidin conjugates. In conclusion, this 125I-pGL2-polylysine-avidin-biotin-T101 monoclonal antibody conjugate can be used to deliver the radioactive DNA to the nuclear fraction of cells.
Citation
Journal of the International Society of Tumor Targeting

Keywords

intracellular localization, luciferase plasmid DNA, T101 monoclonal antibody, transfection

Citation

Chakrabarti, M. , Carrasquillo, J. , Drumm, D. , Neumann, R. and Panyutin, I. (2008), Delivery of DNA Through the CD5 Receptor Using an Avidinbiotin T101 Antibody Conjugate System, Journal of the International Society of Tumor Targeting (Accessed July 25, 2024)

Issues

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Created October 16, 2008