کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5917064 1163769 2014 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Characterization of an anti-Bla g 1 scFv: Epitope mapping and cross-reactivity
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شناسی مولکولی
پیش نمایش صفحه اول مقاله
Characterization of an anti-Bla g 1 scFv: Epitope mapping and cross-reactivity
چکیده انگلیسی


- X-ray crystallography determined the structure of an anti-Bla g 1 scFv.
- The epitope of Bla g 1 was mapped with scanning alanine mutagenesis and ELISA.
- Molecular modeling simulated the complex of the Bla g 1 and the scFv.
- The scFv does not bind Per a 1 from extract nor did it inhibit IgE from one patient.
- The scFv could be used to differentiate Blattella germanica from Periplaneta americana.

Bla g 1 is a major allergen from Blatella germanica and one of the primary allergens used to assess cockroach allergen exposure. The epitope of an anti-Bla g 1 scFv was mapped in order to better understand cross reactivity with other group 1 cockroach allergens and patient IgE epitopes. X-ray crystallography was used to determine the structure of the scFv. The scFv epitope on Bla g 1 was located by alanine scanning site-directed mutagenesis and ELISA. Twenty-six rBla g 1-GST alanine mutants were evaluated for variations in binding to the scFv compared to the wild type allergen. Six mutants showed a significant difference in scFv binding affinity. These mutations clustered to form a discontinuous epitope mainly comprising two helices of Bla g 1. The allergen-scFv complex was modeled based on the results, and the epitope region was found to have low sequence similarity with Per a 1, especially among the residues identified as functionally important for the scFv binding to Bla g 1. Indeed, the scFv failed to bind Per a 1 in American cockroach extract. The scFv was unable to inhibit the binding of IgE antibodies from a highly cockroach allergic patient to Bla g 1. Based on the surface area of Bla g 1 occluded by the scFv, putative regions of patient IgE-Bla g 1 interactions can be inferred. This scFv could be best utilized as a capture antibody in an IgE detection ELISA, or to differentiate Bla g 1 from Per a 1 in environmental exposure assays.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Molecular Immunology - Volume 59, Issue 2, June 2014, Pages 200-207
نویسندگان
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