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Báo cáo khoa học: Catalytic digestion of human tumor necrosis factor-a by antibody heavy chain

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It has long been an important task to prepare a catalytic antibody capable of digesting a targeting crucial protein that controls specific life functions. Tumor necrosis factor-a(TNF-a) is a cytokine and an important molecule concerned with autoimmune diseases such as rheumatoid arthritis, chronic obstructive pulmonary disease, and Crohn’s disease. | ỊFEBS Journal Catalytic digestion of human tumor necrosis factor-a by antibody heavy chain Emi Hifumi1 2 Kyohei Higashi3 and Taizo Uda2 3 1 Research Center for Applied MedicalEngineering Oita University Japan 2 CREST of JST Japan Science and Technology Corporation Kawaguchi Saitama Japan 3 Faculty of Engineering Oita University Japan Keywords catalytic antibody cytokine proteolysis TNF-a Correspondence T. Uda Oita University Faculty of Engineering 700 Dannoharu Oita-shi Oita 870-1192 Japan Fax 81 97 554 7892 Tel 81 97 554 7892 E-mail uda@cc.oita-u.ac.jp Received 10 April 2010 revised 19 June 2010 accepted 21 July 2010 doi 10.1111 j.1742-4658.2010.07785.x It has long been an important task to prepare a catalytic antibody capable of digesting a targeting crucial protein that controls specific life functions. Tumor necrosis factor-a TNF-a is a cytokine and an important molecule concerned with autoimmune diseases such as rheumatoid arthritis chronic obstructive pulmonary disease and Crohn s disease. A mAb ETNF-6 mAb raised against human TNF-a was prepared and the steric conformation was created by using molecular modeling after the cDNA was sequenced. The heavy chain ETNF-6-H of the mAb was considered to possess a catalytic triad-like structure in the complementarity determining regions CDRs . As a result ETNF-6-H exhibited a peptidase and a protease activity. In fact ETNF-6-H predominantly cleaved the Ser5-Arg6 bond of TNF-a at the first step resulting in the generation of a fragment of 17 kDa. This fragment was digested to a smaller molecule of 15 kDa by scission of the Gln21-Ala22 bond. The intermediate product was further converted into a fragment of 13.3 kDa by successive cleavage of the Leu36-Leu37 and Asn39-Gly40 bonds. The heavy chain possessed a protease activity against TNF-a with a multicleavage site. Introduction This decade has seen the preparation of many natural catalytic antibodies. The first natural catalytic antibody isolated from the serum of an .

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