微生物を由来とする機能性タンパク質の医用工学的応用に関する研究

概要

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A study on novel functional protein tool derived from bacterial products was carried out. Thistool is composed of two parts, a functional domain to exhibit regulated cytotoxicy and a targetingdomain to recognize the target cell, and these domains are connected with a chemical linker. APseudomonas aeruginosa exotoxin A (ETA) deficient in its receptor-binding domain, namedΔBD-ETA, was adopted as the functional domain. ΔBD-ETA was expressed in Escherichia coliexpression system and purified by Ni-NTA affinity chromatography. Purified ΔBD-ETA indicates nocytotoxity to human lung carcinoma A549 at the concentrations showing severe cytotoxity ofwild-type ETA. A recombinant single chain Fv (ScFv) derived from anti-carcinoembryonicantigen (CEA) antibody, named anti-CEA-ScFv, was selected as the targeting domain. In contrastto ΔBD-ETA, anti-CEA-ScFv was difficult to express stably as functional targeting domain againstCEA. Thus, the construction of functional protein tool has not been completed at the moment.However, further attempts to overcome the problem including the preparation ofalternative/improved functional protein tool are in progress.