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J.Health Sci., 50(2), 159-163, 2004
Method for Delivering Radiolabeled Single-Chain Fv Antibody to the Brain
Osamu Nakajima,* Akiko Hachisuka, Haruyo Okunuki, Kayoko Takagi, Reiko Teshima, and Jun-ichi Sawada
Division of Biochemistry and Immunochemistry, National
Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku,
Tokyo 158-8501, Japan
Radiolabeled antibody has attractive features as
a therapeutic agent or diagnostic reagent. However, it
is difficult for radiolabeled antibody to enter the
central nervous system (CNS). The purpose of this study
was to develop a method of delivering radiolabeled
single-chain Fv (scFv) antibody into the CNS with the
transactivator of transcription (TAT), one of the
protein transduction domains. Oligonucleotide encoding
TAT was linked to the 5'-terminal of the scFv gene.
The construct was subcloned into the pET-23b vector,
and the recombinant protein was expressed as an
inclusion body in Escherichia coli. After solubilization
and purification, the recombinant protein was
oxidatively labeled with 125I, and the radiolabeled
recombinant protein was injected intraperitoneally into mice.
Six hours later the brains were collected and
homogenized, and the protein fractions were prepared by
acetone precipitation. The radioactivity in the
cerebrum was about 1.6-fold higher in mice administered
TAT-scFv than in those administered scFv alone. The
radiolabeled TAT-scFv antibody was delivered into the
cerebrum more efficiently than radiolabeled scFv
antibody without TAT, suggesting that TAT peptide could
be a candidate tool for delivering radiolabeled scFv
antibody into the CNS.
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