Ricin conjugation using

To prepare an antibody-ricin conjugate using this protocol, 2.25 mg of antibody is needed... 

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. 

Thorpe, P.E., Mason, D.W., Brown, A.N.F., Simmonds, S.J., Ross, W.C.J., Cumber, A.J., and Forrester, J.A. (1982) Selective killing of malignant cells in a leukaemic rat bone marrow using an antibody-ricin conjugate. Nature (London) 297, 594. 

Thorpe PE, Mason DW, Brown AN, et al. Selective killing of malignant cells in leukemic rat bone marrow using an antibody-ricin conjugate. Nature. 1982 297 594-596. 

Conjugations with iodoacetyl crosslinkers have been done using ricin and CVF (Thorpe et al., 1984 Vogel, 1987 Myers et al., 1989). The following generalized protocol for using SIAB is based on the method of Cumber et al. (1985). 

Purification of the immunotoxin conjugate from unconjugated ricin can be done using a column of TSK3000 SW (Toya Soda, Japan) according to the method of Myers et al. (1989). 

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. 

Anti-tumour monoclonal antibodies can also be used to deliver toxins to tumour sites. Toxins conjugated to therapeutic antibodies include ricin, pokeweed toxin, Pseudomonas toxin and... 

The ricin and abnn A chains used to block high affinity binding sites on the column should be alkylated to remove the free sulfhydryl group that could interfere with conjugate purification... 

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