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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Heterologous Expression, Purification and Immunoreactivity of the Antigen 5 from Polybia paulista Wasp Venom

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Autor(es):
Bazon, Murilo Luiz [1] ; Perez-Riverol, Amilcar [1] ; Aparecido dos Santos-Pinto, Jose Roberto [2] ; Romani Fernandes, Luis Gustavo [3] ; Musacchio Lasa, Alexis [4] ; Justo-Jacomini, Debora Lais [1] ; Palma, Mario Sergio [2] ; Zollner, Ricardo de Lima [3] ; Brochetto-Braga, Marcia Regina [1, 5]
Número total de Autores: 9
Afiliação do(s) autor(es):
[1] Univ Estadual Paulista, Lab Biol Mol Artropodes LBMA IB RC UNESP, Av 24-A, 1515, BR-13506900 Rio Claro, SP - Brazil
[2] Univ Estadual Paulista, Ctr Estudos Insetos Sociais CEIS IBRC UNESP, Av 24-A, 1515, BR-13506900 Rio Claro, SP - Brazil
[3] Univ Estadual Campinas UNICAMP, Dept Clin Med, Lab Imunol Translac LIT, Fac Ciencias Med, Rua Tessalia Vieira de Camargo 126, BR-13083887 Campinas, SP - Brazil
[4] Ctr Genet Engn & Biotechnol, Biomed Res Div, Syst Biol Dept, Ave 31, E-158 & 190, POB 6162, Havana 10600 - Cuba
[5] Univ Estadual Paulista, Ctr Estudos Venenos & Animais Peconhentos CEVAP, Fazenda Expt Lageado, Rua Jose Barbosa de Barros 1780, BR-18610307 Botucatu, SP - Brazil
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: TOXINS; v. 9, n. 9 SEP 2017.
Citações Web of Science: 6
Resumo

Polybia paulista (Hymenoptera: Vespidae) is responsible for a high number of sting accidents and anaphylaxis events in Southeast Brazil, Argentina and Paraguay. The specific detection of allergy to the venom of this wasp is often hampered by the lack of recombinant allergens currently available for molecular diagnosis. Antigen 5 (similar to 23 kDa) from P. paulista venom (Poly p 5) is a highly abundant and glycosylated allergenic protein that could be used for development of component-resolved diagnosis (CRD). Here, we describe the cloning and heterologous expression of the antigen 5 (rPoly p 5) from P. paulista venom using the eukaryotic system Pichia pastoris. The expression as a secreted protein yielded high levels of soluble rPoly p 5. The recombinant allergen was further purified to homogeneity (99%) using a two-step chromatographic procedure. Simultaneously, the native form of the allergen (nPoly p 5) was purified from the wasp venom by Ion exchange chromatography. The rPoly p 5 and nPoly p 5 were then submitted to a comparative analysis of IgE-mediated immunodetection using sera from patients previously diagnosed with sensitization to wasp venoms. Both rPoly p 5 and nPoly p 5 were recognized by specific IgE (sIgE) in the sera of the allergic individuals. The high levels of identity found between nPoly p 5 and rPoly p 5 by the alignment of its primary sequences as well as by 3-D models support the results obtained in the immunoblot. Overall, we showed that P. pastoris is a suitable system for production of soluble rPoly p 5 and that the recombinant allergen represents a potential candidate for molecular diagnosis of P.paulista venom allergy. (AU)

Processo FAPESP: 14/13936-7 - Alérgenos de veneno da vespa social Polybia paulista (Hymenoptera, Vespidae): expressão recombinante em leveduras para melhorar o diagnóstico e imunoterapia específica para alergia ao veneno de himenópteros
Beneficiário:Márcia Regina Brochetto Braga
Modalidade de apoio: Auxílio à Pesquisa - Regular
Processo FAPESP: 13/26451-9 - Bioprospecção e Análise Estrutural das Proteínas da Seda de Artrópodes por uma Abordagem Proteômica Utilizando um Sistema nanoLC-ESI-CID/ETD
Beneficiário:José Roberto Aparecido dos Santos-Pinto
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado