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

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  • A Study on Functional Protein Tool Derived from Bacterial Products for Clinical Engineering Application
  • ビセイブツ オ ユライ ト スル キノウセイ タンパクシツ ノ イヨウ コウガクテキ オウヨウ ニ カンスル ケンキュウ

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A study on novel functional protein tool derived from bacterial products was carried out. This tool is composed of two parts, a functional domain to exhibit regulated cytotoxicy and a targeting domain to recognize the target cell, and these domains are connected with a chemical linker. A Pseudomonas aeruginosa exotoxin A (ETA) deficient in its receptor-binding domain, named ΔBD-ETA, was adopted as the functional domain. ΔBD-ETA was expressed in Escherichia coli expression system and purified by Ni-NTA affinity chromatography. Purified ΔBD-ETA indicates no cytotoxity to human lung carcinoma A549 at the concentrations showing severe cytotoxity of wild-type ETA. A recombinant single chain Fv (ScFv) derived from anti-carcinoembryonic antigen (CEA) antibody, named anti-CEA-ScFv, was selected as the targeting domain. In contrast to ΔBD-ETA, anti-CEA-ScFv was difficult to express stably as functional targeting domain against CEA. Thus, the construction of functional protein tool has not been completed at the moment. However, further attempts to overcome the problem including the preparation of alternative/improved functional protein tool are in progress.

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