Ricin A chain conjugation

Hashimoto, N., Takatsu, K., Masuho, Y., Kishida, K., Hara, T., and Hamaoka, T. (1984) Selective elimination of a B cell subset having acceptor site(s) for T cell-replacing factor (TRF) with biotinylated antibody to the acceptor site(s) and avidin-ricin A chain conjugate./. Immunol. 132, 129-135. 

Masuho, Y., Kishida, K., Saito, M., Umemoto, N., and Hara, T. (1982) Importance of the antigen-binding valency and the nature of the cross-linking bond in ricin A-chain conjugates with antibody. J. Biochem. (Tokyo) 91, 1583. 

Raso, V., and Basala, M. (1984) A highly cytotoxic human transferrin - Ricin A chain conjugate used to select receptor-modified cells./. Biol. Chem. 259, 1143. 

Raso V, Ritz J, Basala M, et al. Monoclonal antibody-ricin A chain conjugate selectively cytotoxic for cells bearing the common acute lymphoblastic leukemia antigen. Cancer Res 1982 42 457-464. 

Pincus, S. H., Wehrly, K., and Chesebro, B. (1989) Treatment of HIV tissue culture infection with monoclonal antibody-ricin A chain conjugates. J. Immunol. 142, 3070-3075. 

Raso, V, and Basala, M. (1985) Study of the transferrin receptor using a cytotoxic human transferrin—ricin A chain conjugate, in Receptor-Mediated Targeting of Drugs (G. Gre-goriadis, ed.), Vol. 2, p. 73. Plenum, New York. 

Ghetie, V., Till, M.A., Ghetie, M.-A., Tucker, T., Porter, J., Patzer, E.J., Richardson, J.A., Uhr, J.W., and Vitetta, E.S. (1990) Preparation and characterization of conjugates of recombinant CD4 and deglyco-sylated ricin A chain using different crosslinkers. Bioconjugate Chem. 1, 24-31. 

Antibody-toxin conjugates made with ricin A chain, abrin A chain, gelomn, and momordin can be stored for at least 4 yr at -70°C without detectable loss of activity. The bond between the antibody and the RIP breaks down very slowly at 4°C in PBSE but, provided that care is taken to ensure the sterility of the solution, conjugates can be stored under these conditions for up to one year with little deterioration in quality. 

RIPs should generally appear as a single band on SDS-PAGE. The presence of additional bands is a clear indication of impurity, and may complicate the interpretation of electrophoretic analysis of conjugate products. The exception is ricin A chain that has been isolated from the native toxin. Native ricin A chain exists as a mixture of two differently... 

The capacity of immunoaffinity HPLC columns prepared using affinity-purified antibody against ricin A chain, abrin A chain, gelonin, and momordin varies between 400 and 600 pg/mL of bed vol for the appropriate RIP. In the case of the antibody-toxin conjugates made with these RIPs, the capacity is at least 2 mg/mL of bed vol. 

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