A novel monoclonal antibody against the c-terminal region of aquaporin-4

Julia Ramadhanti, Ping Huang, Osamu Kusano-Arai, Hiroko Iwanari, Toshiko Sakihama, Tasturo Misu, Kazuo Fujihara, Takao Hamakubo, Masato Yasui, Yoichiro Abe

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

Aquaporin-4 (AQP4), the most abundant water channel in the brain, plays a central role in water homeostasis, neuronal activity, and migration of astrocytes in the central nervous system. Recent studies have demonstrated that AQP4 is a target of an autoantibody specifically detected in an autoimmune neurologic disease called neuromyelitis optica. Here we have generated a monoclonal antibody (MAb) against the C-terminal region of AQP4 using a baculovirus expressing mouse AQP4 as an immunogen. This antibody (clone E5206) recognized both human and mouse AQP4s in a denaturing condition and was able to precipitate AQP4 from cell lysates of CHO cells stably expressing AQP4. Western blot analysis using deletion mutants revealed that the epitope was located within a region between Asp303 and Leu320 in the C-terminal tail of AQP4. Although clone E5206 could not be used for immunostaining when cells or tissues were fixed with 4% paraformaldehyde or 10% formalin, it could be used when cells were fixed with 10% trichloroacetic acid and when a formalin-fixed tissue section was pretreated with antigen-retrieval reagents. This MAb can be a valuable tool for analysis of AQP4 in a variety of physiological and pathophysiological contexts, in human tissues and organs as well as in rodent models, both in vitro and in vivo.

Original languageEnglish
Pages (from-to)270-276
Number of pages7
JournalMonoclonal Antibodies in Immunodiagnosis and Immunotherapy
Volume32
Issue number4
DOIs
Publication statusPublished - 2013 Aug 1

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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