Establishment of a novel anti-TROP2 monoclonal antibody TrMab-29 for immunohistochemical analysis

Yusuke Sayama, Mika K. Kaneko, Junko Takei, Hideki Hosono, Masato Sano, Teizo Asano, Yukinari Kato

Research output: Contribution to journalArticlepeer-review

Abstract

TROP2 is a type I transmembrane glycoprotein originally identified in human trophoblast cells that is overexpressed in several types of cancer. To better understand the role of TROP2 in cancer, we herein aimed to develop a sensitive and specific anti-TROP2 monoclonal antibody (mAb) for use in flow cytometry, Western blot, and immunohistochemistry using a Cell-Based Immunization and Screening (CBIS) method. Two mice were immunized with N-terminal PA-tagged and C-terminal RAP/MAP-tagged TROP2-overexpressed Chinese hamster ovary (CHO)–K1 cells (CHO/PA-TROP2-RAP-MAP), and hybridomas showing strong signals from PA-tagged TROP2-overexpressed CHO–K1 cells (CHO/TROP2-PA) and weak-to-no signals from CHO–K1 cells were selected using flow cytometry. We demonstrated using flow cytometry that the established anti-TROP2 mAb, TrMab-29 (mouse IgG1 kappa), detected TROP2 in MCF7 breast cancer cell line as well as CHO/TROP2-PA cells. Western blot analysis showed a 40 kDa band in lysates prepared from both CHO/TROP2-PA and MCF7 cells. Furthermore, TROP2 was strongly detected by immunohistochemical analysis using TrMab-29, indicating that TrMab-29 may be a valuable tool for the detection of TROP2 in cancer.

Original languageEnglish
Article number100902
JournalBiochemistry and Biophysics Reports
Volume25
DOIs
Publication statusPublished - 2021 Mar

Keywords

  • Breast cancer
  • CBIS method
  • Monoclonal antibody
  • TROP2

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

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