Establishment of an Anti-CD20 Monoclonal Antibody (C20Mab-60) for Immunohistochemical Analyses

Yoshikazu Furusawa, Mika K. Kaneko, Yukinari Kato

Research output: Contribution to journalArticlepeer-review

Abstract

Sensitive and specific monoclonal antibodies (mAbs) are needed for detecting CD20. This antigen, one of several B lymphocyte antigens, is a target for every application used in the diagnosis of B cell lymphoma. Many anti-CD20 mAbs have been established, although applications of these antibodies are limited. This study aims to establish sensitive and specific anti-CD20 mAbs suitable for broad application, such as flow cytometry, Western blotting, and immunohistochemical analyses. Using the Cell-Based Immunization and Screening (CBIS) method, all procedures were performed by utilizing CD20-stable transfectants, and a clone, C20Mab-60 (IgG2a, kappa), was developed. In flow cytometry, C20Mab-60 detected overexpression of CD20 in LN229 cell and endogenous CD20 in BALL-1 (a human B cell leukemia cell line) but did not react with CD20-knockout BALL-1 (BINDS-24), indicating specificity for CD20. In Western blotting, C20Mab-60 detected CD20-overexpressing Chinese hamster ovary-K1, BALL-1, and Raji (a human Burkitt's lymphoma cell line) displaying both sensitivity and specificity. Furthermore, B cell but not T cell lymphomas were strongly stained with C20Mab-60 in immunohistochemical analyses. C20Mab-60, which was developed by CBIS method, is shown to be useful for the detection of cells expressing CD20 in lymphoma tissues by flow cytometry, Western blotting, and immunohistochemical analyses.

Original languageEnglish
Pages (from-to)112-116
Number of pages5
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume39
Issue number4
DOIs
Publication statusPublished - 2020 Aug 1

Keywords

  • CD20
  • Western blotting
  • flow cytometry
  • immunohistochemistry
  • monoclonal antibody

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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