کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
9902278 1545797 2005 11 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Production of IgA monoclonal antibody against Shiga toxin binding subunits employing nasal-associated lymphoid tissue
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Production of IgA monoclonal antibody against Shiga toxin binding subunits employing nasal-associated lymphoid tissue
چکیده انگلیسی
We established an IgA monoclonal antibody (mAb) against Shiga toxin 1 B subunits (Stx1B) from mouse nasal-associated lymphoid tissues (NALT) of BALB/c mice. We have developed an improved protocol in which cross-linked Stx1B is intranasally administered together with cholera toxin. Surface IgA-positive NALT lymphocytes from mice immunized in this manner were enriched and then fused with mouse myeloma cells to produce hybridoma cells. Hybridoma culture supernatants were examined to see if they contain IgA against Stx1B and if they can inhibit carbohydrate recognition by Stx1B. For the latter purpose, we prepared carbohydrate ligands in which globotriose is present on the poly-lysine backbone. The established IgA mAb exhibited saturable and dose-dependent binding to the immobilized Stx1B. Inversely, the binding of the carbohydrate ligands to the immobilized Stx1B was inhibited by the mAb pretreatment. Immunoblotting and SDS-PAGE analysis revealed dimeric IgA. The IgA mAb inhibited the binding of digoxigenin-conjugated Stx1B to natural ligands displayed on a Burkitt's lymphoma cell line, Ramos. These results suggested that surface IgA-positive B cells in the inductive sites of the mucosal immune system in the upper respiratory tract are a potent source for producing IgA mAb against protein antigens with weak immunogenicity such as Stx1B.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 302, Issues 1–2, July 2005, Pages 125-135
نویسندگان
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