Research Papers:

Antitumor activity of a newly developed monoclonal antibody against ROR1 in ovarian cancer cells

Zhengna Yin, Mengyun Gao, Sasa Chu, Yiping Su, Chunping Ye, Yiquan Wang, Zhuanqin Pan, Zhuming Wang, Huilin Zhang, Hua Tong _ and Jin Zhu

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Oncotarget. 2017; 8:94210-94222. https://doi.org/10.18632/oncotarget.21618

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Zhengna Yin1,*, Mengyun Gao1,*, Sasa Chu2, Yiping Su1, Chunping Ye1, Yiquan Wang3, Zhuanqin Pan4, Zhuming Wang5, Huilin Zhang1, Hua Tong1 and Jin Zhu6

1Department of Obstetrics and Gynecology, Nanjing Maternity and Child Health Care Hospital, Obstetrics and Gynecology Hospital Affiliated to Nanjing Medical University, Nanjing 210004, China

2Department of Infectious Disease, Institute of Liver Disease, Nanjing Jingdu Hospital, Nanjing 210002, China

3Department of Traditional Chinese Internal Medicine, Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200071, China

4Department of Nursing, Gaoyou People’s Hospital, Yangzhou 225600, China

5Department of Pathology, Chinese Ministry of Health-designated Key Laboratory of Antibody Technology, Nanjing Medical University, Nanjing 210029, China

6Huadong Medical Institute of Biotechniques, Nanjing 210002, China

*These authors have contributed equally to this work

Correspondence to:

Hua Tong, email: [email protected]

Huilin Zhang, email: [email protected]

Keywords: ROR1, monoclonal antibody production, chimeric antibody Fab, binding affinity, antitumor activity

Received: July 12, 2017    Accepted: August 29, 2017    Published: October 07, 2017


Receptor-tyrosine-kinase-like Orphan Receptor 1 (ROR1) is a tyrosine-protein kinase transmembrane receptor and ROR1 overexpression is associated with a poor prognosis in various cancers, including ovarian cancer. Targeting of ROR1 has been evaluated as a novel cancer therapy strategy. This study developed a novel chimeric anti-ROR1 Fab antibody (named ROR1-cFab) and then assessed the antitumor activity of this antibody in ovarian cancer cells, an in vitro model of preclinical cancer therapy. A ROR1-cFab prokaryotic expression vector was constructed from positive fusion cells (splenocytes from mice with high ROR1 immune titers were fused with myeloma cells) after three rounds of sub-clone affinity screening. Then, a variety of assays were employed to assess the binding selectivity and specificity of ROR1-cFab to ROR1 protein. Furthermore, CCK8, flow cytometric apoptosis, wound healing, and Transwell migration assays were used to assess antitumor activity of this newly developed anti-ROR1 antibody in ovarian cancer cells. We demonstrated that ROR1-cFab could specifically bind to ROR1 protein and ROR1-positive ovarian cancer A2780 cells. Functional assays revealed that ROR1-cFab inhibited tumor cell proliferation and migration, as well as inducing apoptosis of ROR1-positive A2780 cells in a dose dependent manner. These effects were not observed in ROR1-negative lose386 cells. In conclusion, ROR1-cFab is a novel anti-ROR1 antibody with a high affinity to ROR1 protein and inhibitory effects on ROR1-positive cells. Future studies will determine whether the ROR1-cFab might be a promising candidate for treatment of ROR1-positive ovarian cancer.

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