Epitope analysis of peanut allergen Ara h1 with human monoclonal IgM antibody clone #86

Article type: Research Article

Authors: Shinmoto, Hiroshi^{a; *} | Takeda, Mio^a | Matsuo, Yuji^a | Naganawa, Yasunori^b | Tomita, Shinichi^a | Takano-Ishikawa, Yuko^c

Affiliations: [a] Tamagawa University, Machida, Tokyo, Japan | [b] Nagahama Institute of Bio-Science and Technology, Nagahama, Japan | [c] National Food Research Institute, Tsukuba, Japan

Correspondence: [*] Corresponding author: Hiroshi Shinmoto, Ph.D., Tamagawa University, Tamagawa-gakuen 6-1-1, Machida, Tokyo, 194-8610 Japan. Tel./Fax: +81 42 739 8267; E-mail: [email protected].

Abstract: A human-mouse hybridoma clone #86 secreting IgM-class human monoclonal antibody to peanut allergen protein Ara h1 was newly established. To detect an antibody-binding sequence (epitope) on Ara h1, the monoclonal antibody #86 was reacted with multi-pin apparatus with a series of peptides synthesized from the amino acid sequence of Ara h1. The antibody #86 was found to bind to a peptide with amino acid sequence of 481EEEEDEDEEEEGSNREVRRY500. Further analysis with shorter pin-peptides with ten amino acid-long showed that the peptides reacted with the antibody #86 contained a sequence of 485DEDEEEE491. This might be an essential linear sequence of this epitope. When the 485DED487 part of the peptide was replaced by alanine, decreased binding of antibody #86 was observed.

Keywords: Human monoclonal antibody, allergen, peanut, epitope, IgM

DOI: 10.3233/HAB-2010-0233

Journal: Human Antibodies, vol. 19, no. 4, pp. 101-105, 2010