SpringerProtocols: Abstract: 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

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

By: Sun U. Song, Wayne A. Marasco

Abstract

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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.

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Fig. 1. Schematic diagram of Fab(Tac)-protamine fusion protein. Recombinant Fab(Tac) antibody is composed of Fd fragment (V_H and C_H1), and κchain (V_L and C_L) which are connected by a disulfide bond. Protamine protein is fused with the carboxyl end of Fd.

Book Title: Gene Targeting Protocols

Series: Methods in Molecular Biology | Volume: 133 | Pub. Date: Sep-23-1999 | Page Range: 157-166 | DOI: 10.1385/1-59259-215-5:157

Subject: Genetics/Genomics

References

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