Preparation and Cytotoxic Potency of Rabbit Immunoglobulin Fab'-Ricin A-chain Hybrid Molecules-A Technical Note

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概要

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• As an approach to the development of new antitumor agents, we described procedures for the preparation of a new agent, i. e., antimelanoma antibody (Fab') conjugated with ricin A-chain which is a subunit of a highly toxic plant toxin, and studied its effect on melanoma cells in vitro. Antibodies to human malignant melanoma (KH_<m-1> cell line) were produced in rabbits by inoculating the cells suspended in Freund's complete adjuvant, and the Fab' fragments purified from isolated immunoglobulin G (IgG). Ricin A-chain was prepared from ricin isolated from Ricinus communis beans. The conjugates of Fab' and ricin A-chain were prepared through a disulfide bond and isolated by column chromatography. The hybrid molecules exhibited complete cytotoxicity to KH_<m-1> melanoma cells in vitro, at a concentration of 6×10^<-8> M. In contrast, none of unconjugated Fab' or ricin A-chain alone, their mixtures, and the hybrid made of the Fab' fragment of normal IgG and ricin A-chain showed any significant cytotoxicity even at higher concentrations (1×10^<-6>, 1.4×10^<-6>, 1×10^<-6> and 5.4×10^<-7> M respectively). A mixture of anti-KH_<m-1> IgG and rabbit complement showed some cytotoxicity to KH_<m-1> melanoma cells but the potency was 1/100 of that of the antibody Fab'-Ricin A-chain hybrid. These data suggest that the resulting hybrid exerts the toxic effect of ricin A-chain on melanoma cells through binding of its Fab' moiety to cell surface antigens.

• 北里大学の論文
• 1983-06-30

著者

• Kanzaki Tamotsu

  Department of Dermatology, Kagoshima University Faculty of Medicine

• Kanzaki Tamotsu

  Department Of Dermatology Kitasato University School Of Medicine

• Kanamaru Tetsuzan

  Department of Dermatology, Kitasato University School of Medicine

• Masuho Yasuhiko

  Department of Cancer Therapeutics

• Hamazaki Hideaki

  Department of Biochemistry, Kitasato University School of Medicine

• Kanzaki Tamotsu

  Department Of Dermatology Faculty Of Medicine Kagoshima University

• Hamazaki Hideaki

  Department Of Biochemistry Kitasato University School Of Medicine

• Kanamaru Tetsuzan

  Department Of Dermatology Kitasato University School Of Medicine

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