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PLoS One. 2014 Dec 1;9(12):e113442. doi: 10.1371/journal.pone.0113442. eCollection 2014.

Effective therapeutic approach for head and neck cancer by an engineered minibody targeting the EGFR receptor.

Author information

1
Department of Bio-Engineering, Life Science RD Center, Sinil Pharmaceutical Co., Seongnam, South Korea.
2
Department of Physiology, Ajou University School of Medicine, Suwon, South Korea.
3
Department of Physiology, Ajou University School of Medicine, Suwon, South Korea; Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea.
4
Department of Neuroscience, Kyung Hee University School of Medicine, Seoul, South Korea.
5
Division of Pharmacology, Department of Molecular Cell Biology, Samsung Biomedical Research Institute, SungKyunKwan University School of Medicine, Suwon, South Korea.
6
Well Aging Research Center, Samsung Advanced Institute of Technology (SAIT), Suwon, South Korea.
7
Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea; Department of Biochemistry, Ajou University School of Medicine, Suwon, South Korea.

Abstract

Cetuximab, a chimeric monoclonal antibody developed for targeting the Epidermal Growth Factor Receptor (EGFR), has been intensively used to treat cancer patients with metastatic colorectal cancer and head and neck cancer. Intact immunoglobulin G (IgG) antibody like cetuximab, however, has some limitations such as high production cost and low penetration rate from vasculature into solid tumor mass due to its large size. In attempt to overcome these limitations, we engineered cetuximab to create single chain variable fragments (scFv-CH3; Minibody) that were expressed in bacterial system. Among three engineered minibodies, we found that MI061 minibody, which is composed of the variable heavy (VH) and light (VL) region joined by an 18-residue peptide linker, displays higher solubility and better extraction properties from bacterial lysate. In addition, we validated that purified MI061 significantly interferes ligand binding to EGFR and blocks EGFR's phosphorylation. By using a protein microarray composed of 16,368 unique human proteins covering around 2,400 plasma membrane associated proteins such as receptors and channels, we also demonstrated that MI061 only recognizes the EGFR but not other proteins as compared with cetuximab. These results indicated that engineered MI061 retains both binding specificity and affinity of cetuximab for EGFR. Although it had relatively short half-life in serum, it was shown to be highly significant anti-tumor effect by inhibiting ERK pathway in A431 xenograft model. Taken together, our present study provides compelling evidence that engineered minibody is more effective and promising agent for in vivo targeting of solid tumors.

PMID:
25438047
PMCID:
PMC4249956
DOI:
10.1371/journal.pone.0113442
[Indexed for MEDLINE]
Free PMC Article

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