Epitope analysis of peanut allergen Ara h1 with oligoclonal IgM antibody from human B-lymphoblastoid cells

Abstract

To analyze epitopes of peanut allergen Ara h1, Epstein-Barr virus-transformed human peripheral oligoclonal B-cells were cultured to obtain antibodies to Ara h1. The combined reaction pattern with six oligoclonal antibodies showed there were six antibody binding areas named a to f in Ara h1. We found the novel antibody binding area named "area c" (171-230aa).

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

“…Peripheral blood lymphocytes from a healthy donor were transformed by Epstein-Barr virus infection [8,9,12]. The resulted human B-lymphoblastoid cells secreting IgM-class antibody to Ara h1 were fused with mouse myeloma SP2/O3 cells with 50% polyethylenglycol.…”

“…Several peanut proteins have been identified as allergens by using sera from patients allergic to peanut, and Ara h1, Ara h2, Ara h3, and Ara h4 have been reported as major peanut allergens [3][4][5][6]. In the previous report, we established a mouse-human hybridoma clone #92-2 secreting IgM-class human monoclonal antibodies to Ara h1 from peripheral blood lymphocytes [7][8][9], and showed novel linear B-cell epitope of Ara h1. In this study, an another clone of human-mouse hybridoma (clone #86) secreting human IgM class monoclonal antibody to Ara h1 was established for further epitope analysis of Ara h1.…”

Epitope analysis of peanut allergen Ara h1 with human monoclonal IgM antibody 92-2

“…Several peanut proteins have been identified as allergens by using sera from patients allergic to peanut, and Ara h1 and Ara h2 have been reported as major peanut allergens (de Jong et al 1998;Koppelman et al 2004;Burks et al 1991, Maleki et al 2000. We have established five clones of mouse-human hybridomas secreting IgM-class human monoclonal antibodies to peanut allergens from peripheral blood lymphocytes transformed with Epstein-Barr virus followed by cell fusion with mouse myeloma cells (Naganawa et al 2008;Shinmoto et al 2004). In this paper, we present the results of a linear B-cell epitope analysis with synthesized Ara h1 oligo peptides recognized by IgM-class human monoclonal antibody 92-2.…”