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  • Title: Expression of ricin A chain and ricin A chain-KDEL in Escherichia coli.
    Author: Zhan J, Chen Y, Wang K, Zheng S.
    Journal: Protein Expr Purif; 2004 Apr; 34(2):197-201. PubMed ID: 15003251.
    Abstract:
    Ricin and its A chains can be used to conjugate with monoclonal antibodies to prepare immunotoxins. Ricin A chain (RTA) and its modification RTA-KDEL (ER-retrieval signal) were expressed with the pKK223.3 system in Escherichia coli under control of a tac promoter. The recombinant proteins can be purified by one-step affinity chromatography on a column of Blue-Sepharose 6B. The toxicities of RTA and its mutant RTA-KDEL were evaluated by the MTT assay in HeLa, MCF, and ECV-304 cells following fluid-phase endocytosis. RTA-KDEL was somewhat more cytotoxic than RTA itself in the different cell lines. The results suggest that rRTA-KDEL may be useful for the synthesis of more potent immunotoxins.
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