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KMID : 1094720160210020321
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Biotechnology and Bioprocess Engineering
2016 Volume.21 No. 2 p.321 ~ p.330
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Periplasmic expression, purification, and characterization of an anti-epidermal growth factor receptor antibody fragment in Escherichia coli
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Chi Won-Jae
Kim Hye-Rim
Yoo Hee-Jung
Kim Young-Pil
Hong Soon-Kwang
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Abstract
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New structural designs of antibody fragments have considerable biotechnological and therapeutic potential. In this study, we describe the construction and functional expression of a cetuximab-based antibody fragment (scFv-CH3, minibody) that exhibits activity against human colon cancer. Heterologous expression in Escherichia coli (E. coli) was improved by optimizing the host cells, signal peptides, induction conditions, and culture media. The recombinant minibody was expressed successfully in the periplasm of E. coli BL21(DE3) and purified by immobilized metal affinity chromatography using a Ni2+-NTA resin. The purified minibody showed high binding affinity to cell-surface epidermal growth factor receptor (EGFR) and exhibited inhibition of EGFR-mediated signal transduction in the human colon cancer cell line HT29 in a similar way by the cetuximab. The minibody also showed significant level of anti-cancer ability in the HT29 colorectal cancer xenograft model, which was lower than that by the cetuximab.
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KEYWORD
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minibody, anti-EGFR antibody, colon cancer, anti-cancer activity
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