Abstract：
: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.
journal_name
Mol Immunoljournal_title
Molecular immunologyauthors
Mueller GA,Ankney JA,Glesner J,Khurana T,Edwards LL,Pedersen LC,Perera L,Slater JE,Pomés A,London REdoi
10.1016/j.molimm.2014.02.003subject
Has Abstractpub_date
2014-06-01 00:00:00pages
200-7issue
2eissn
0161-5890issn
1872-9142pii
S0161-5890(14)00030-3journal_volume
59pub_type
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更新日期：2008-06-01 00:00:00
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pub_type： 杂志文章
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更新日期：2005-02-01 00:00:00
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更新日期：2015-03-01 00:00:00
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更新日期：2020-04-01 00:00:00
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pub_type： 杂志文章
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更新日期：1997-08-01 00:00:00
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pub_type： 杂志文章,评审
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更新日期：2004-06-01 00:00:00
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pub_type： 杂志文章
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更新日期：1982-12-01 00:00:00
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pub_type： 杂志文章
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更新日期：2010-09-01 00:00:00
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pub_type： 杂志文章
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更新日期：1983-01-01 00:00:00
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pub_type： 历史文章,杂志文章,评审
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更新日期：2020-03-01 00:00:00
