4OUO

anti-Bla g 1 scFv

Summary for 4OUO

• Entry DOI: 10.2210/pdb4ouo/pdb
• Descriptor: anti Bla g 1 scFv, SULFATE ION, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: cockroach allergen, igg, scfv antibody fragment, bla g 1, immune system
• Biological source: Gallus gallus
• Total number of polymer chains: 1
• Total formula weight: 24823.40

Authors

Mueller, G.A.,Ankney, J.A.,Glesner, J.,Khurana, T.,Edwards, L.L.,Pedersen, L.C.,Perera, L.,Slater, J.E.,Pomes, A.,London, R.E. (deposition date: 2014-02-18, release date: 2014-03-05, Last modification date: 2023-09-20)

Primary citation

Mueller, G.A.,Ankney, J.A.,Glesner, J.,Khurana, T.,Edwards, L.L.,Pedersen, L.C.,Perera, L.,Slater, J.E.,Pomes, A.,London, R.E.
Characterization of an anti-Bla g 1 scFv: Epitope mapping and cross-reactivity.
Mol.Immunol., 59:200-207, 2014

PubMed 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.

PubMed: 24667070
DOI: 10.1016/j.molimm.2014.02.003

Experimental method

X-RAY DIFFRACTION (1.8 Å)