کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3418379 1225511 2008 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Ani s 1, the major allergen of Anisakis simplex: Purification by affinity chromatography and functional expression in Escherichia coli
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی انگل شناسی
پیش نمایش صفحه اول مقاله
Ani s 1, the major allergen of Anisakis simplex: Purification by affinity chromatography and functional expression in Escherichia coli
چکیده انگلیسی

Third stage larvae of the nematode Anisakis simplex often infect marine fish and invertebrates. When the larvae are ingested orally via seafood, they can cause IgE-mediated allergic reactions as well as anisakiasis. Of the known A. simplex allergens, Ani s 1 (Kunitz/bovine pancreatic trypsin inhibitor family protein) has been demonstrated to be a major allergen, being expected to be a useful tool for diagnosis of A. simplex allergy. For a diagnostic purpose, sufficient amounts of either natural Ani s 1 (nAni s 1) or recombinant Ani s 1 (rAni s 1) with an IgE-binding capacity should be stably supplied whenever needed. In this study, therefore, we first developed a simple and rapid purification method for Ani s 1 that is based on affinity chromatography using anti-Ani s 1 antibodies as ligands. The method was shown to produce nAni s 1 with a higher yield than the previously reported methods. Then, an attempt was made to express rAni s 1 in Escherichia coli as a His-tagged protein. rAni s 1 obtained as an inclusion body was solubilized in a solvent containing denaturing and reducing reagents and purified by nickel-chelate chromatography. Refolding of rAni s 1 was accomplished by dialysis in the presence of arginine, followed by that in the absence of arginine. Fluorescence ELISA and inhibition ELISA data revealed that rAni s 1 is IgE reactive enough to be used as a diagnostic tool.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Parasitology International - Volume 57, Issue 3, September 2008, Pages 314–319
نویسندگان
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