Next Article in Journal
Orally Delivered Scorpion Antimicrobial Peptides Exhibit Activity against Pea Aphid (Acyrthosiphon pisum) and Its Bacterial Symbionts
Previous Article in Journal
Botulinum Toxin for the Treatment of Neuropathic Pain
Article Menu
Issue 9 (September) cover image

Export Article

Open AccessArticle
Toxins 2017, 9(9), 259; https://doi.org/10.3390/toxins9090259

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

1
Laboratório de Biologia Molecular de Artrópodes-LBMA-IB-RC-UNESP, Univ Estadual Paulista, Av. 24-A, n° 1515, Bela Vista, Rio Claro, SP CEP 13506-900, Brazil
2
Centro de Estudos de Insetos Sociais-CEIS-IBRC-UNESP, Univ Estadual Paulista, Av. 24-A, n° 1515, Bela Vista, Rio Claro, SP CEP 13506-900, Brazil
3
Laboratório de Imunologia Translacional-LIT, Departamento de Clínica Médica, Faculdade de Ciências Médicas, FCM, Universidade Estadual de Campinas-UNICAMP, Rua Tessália Vieira de Camargo n° 126, Cidade Universitária “Zeferino Vaz”, Campinas, SP CEP 13083-887, Brazil
4
System Biology Department, Biomedical Research Division, Center for Genetic Engineering and Biotechnology, Ave. 31, e/ 158 and 190, P.O. Box 6162, Cubanacan, Playa, Havana 10600, Cuba
5
Centro de Estudos de Venenos e Animais Peçonhentos-CEVAP, Univ Estadual Paulista, Rua José Barbosa de Barros, 1780, Fazenda Experimental Lageado, Botucatu, SP CEP 18610-307, Brazil
*
Author to whom correspondence should be addressed.
Academic Editor: Irina Vetter
Received: 25 July 2017 / Revised: 15 August 2017 / Accepted: 18 August 2017 / Published: 24 August 2017
(This article belongs to the Section Animal Venoms)
View Full-Text   |   Download PDF [4504 KB, uploaded 24 August 2017]   |  

Abstract

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 (~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. View Full-Text
Keywords: Polybia paulista; antigen 5; heterologous expression; allergy; diagnosis Polybia paulista; antigen 5; heterologous expression; allergy; diagnosis
Figures

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Share & Cite This Article

MDPI and ACS Style

Bazon, M.L.; Perez-Riverol, A.; dos Santos-Pinto, J.R.A.; Fernandes, L.G.R.; Lasa, A.M.; Justo-Jacomini, D.L.; Palma, M.S.; Zollner, R.L.; Brochetto-Braga, M.R. Heterologous Expression, Purification and Immunoreactivity of the Antigen 5 from Polybia paulista Wasp Venom. Toxins 2017, 9, 259.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Toxins EISSN 2072-6651 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top