Abstract
A recombinant bifunctional fusion protein, consisting of a recombinant antibody and a DNA-binding protein, can be used as a nonviral gene delivery vector (Fig. 1). In this lab, such a fusion protein, composed of a human antibody Fab(105) moiety against the envelope glycoprotein of HIV-1, gp120, and a human DNA-binding moiety (protamine), was developed (1). Chen et al. (1) showed that the bifunctional fusion protein complexed with plasmid DNA, encoding the catalytic subunit of Pseudomonas exotoxin, a complex known as a genetic immunotoxin, and was specifically transferred into HIV-1-infected cells by receptor-mediated endocytosis, resulting in selective killing of the target cells. However, the low level of Fab(105)-protamine fusion protein secreted from the stably transduced COS cells has been a limiting factor for experiments requiring large amounts of the fusion protein.
Schematic diagram of Fab(Tac)-protamine fusion protein. Recombinant Fab(Tac) antibody is composed of Fd fragment (VH and CH1), and κchain (VL and CL) which are connected by a disulfide bond. Protamine protein is fused with the carboxyl end of Fd.
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Song, S.U., Marasco, W.A. (2000). Rapid Establishment of Myeloma Cell Lines Expressing Fab(Tac)-Protamine, a Targetable Protein Vector, Directed Against High-Affinity α-Chain of Human Interleukin-2 Receptor. In: Kmiec, E.B. (eds) Gene Targeting Protocols. Methods in Molecular Biology™, vol 133. Humana Press. https://doi.org/10.1385/1-59259-215-5:157
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DOI: https://doi.org/10.1385/1-59259-215-5:157
Publisher Name: Humana Press
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Online ISBN: 978-1-59259-215-9
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