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Monoclon Antib Immunodiagn Immunother. 2017 Aug;36(4):143-148. doi: 10.1089/mab.2017.0026. Epub 2017 Jul 12.

H2Mab-77 is a Sensitive and Specific Anti-HER2 Monoclonal Antibody Against Breast Cancer.

Author information

1
1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine , Aoba-ku, Sendai, Miyagi, Japan .
2
2 Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University , Yushima, Bunkyo-ku, Tokyo, Japan .
3
3 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Aoba-ku, Sendai, Miyagi, Japan .
4
4 Department of Pathology and Laboratory Medicine, Sendai Medical Center , Miyagino, Miyagino-ku, Sendai, Miyagi, Japan .
5
5 New Industry Creation Hatchery Center, Tohoku University , Aoba-ku, Sendai, Miyagi, Japan .

Abstract

Human epidermal growth factor receptor 2 (HER2) plays a critical role in the progression of breast cancers, and HER2 overexpression is associated with poor clinical outcomes. Trastuzumab is an anti-HER2 humanized antibody that leads to significant survival benefits in patients with HER2-positive metastatic breast cancers. In this study, we developed novel anti-HER2 monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. Initially, we expressed the full length or ectodomain of HER2 in LN229 glioblastoma cells and then immunized mice with ectodomain of HER2 or LN229/HER2, and performed the first screening by enzyme-linked immunosorbent assays using ectodomain of HER2. Subsequently, we selected mAbs according to their efficacy in flow cytometry (second screening), Western blot (third screening), and immunohistochemical analyses (fourth screening). Among 100 mAb clones, only three mAbs reacted with HER2 in Western blot, and clone H2Mab-77 (IgG1, kappa) was selected. Finally, immunohistochemical analyses with H2Mab-77 showed sensitive and specific reactions against breast cancer cells, warranting the use of H2Mab-77 to detect HER2 in pathological analyses of breast cancers.

KEYWORDS:

HER2; breast cancer; immunohistochemistry; monoclonal antibody

PMID:
28700270
DOI:
10.1089/mab.2017.0026
[Indexed for MEDLINE]

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