Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/22808
Title: Heparin removal by ecteola-cellulose pre-treatment enables the use of plasma samples for accurate measurement of anti-Yellow fever virus neutralizing antibodies
Authors: Azevedo, Ana Carolina Campi
Pascoal, Vanessa Peruhype Magalhães
Reis, Jordana Grazziela Coelho dos
Pereira, Christiane Costa
Yamamura, Anna Yoshida
Lima, Sheila Maria Barbosa de
Simões, Marisol
Campos, Fernanda Magalhães Freire
Tonini, Aline de Castro Zacche
Lemos, Elenice Moreira
Brum, Ricardo Cristiano
Noronha, Tatiana Guimarães de
Freire, Marcos Silva
Maia, Maria de Lourdes Sousa
Camacho, Luiz Antônio Bastos
Rios, Maria
Chancey, Caren
Romano, Alessandro
Domingues, Carla Magda
Carvalho, Andréa Teixeira de
Martins Filho, Olindo Assis
Affilliation: Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brazil
Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brazil
Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brazil
Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brazil
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brazil
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brazil
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brazil
Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brazil
Universidade Federal do Espírito Santo. Núcleo de Doenças Infecto-Parasitárias. Espírito Santo, Brazil
Universidade Federal do Espírito Santo. Núcleo de Doenças Infecto-Parasitárias. Espírito Santo, Brazil
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Assessoria Clínica. Rio de Janeiro, RJ, Brazil
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Assessoria Clínica. Rio de Janeiro, RJ, Brazil
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brazil
Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos. Assessoria Clínica. Rio de Janeiro, RJ, Brazil
Fundação Oswaldo Cruz. Escola Nacional de Saúde Pública.Rio de Janeiro, RJ, Brazil
US Food and Drug Administration. Center for Biologics Evaluation and Research-CBER. Silver Spring, MD, USA
US Food and Drug Administration. Center for Biologics Evaluation and Research-CBER. Silver Spring, MD, USA
Ministério da Saúde. Secretaria de Vigilância em Saúde. Brasília, DF, Brazil
Ministério da Saúde. Secretaria de Vigilância em Saúde. Brasília, DF, Brazil
Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brazil
Fundação Oswaldo Cruz. Instituto René Rachou. Grupo Integrado de Pesquisas em Biomarcadores. Belo Horizonte, MG, Brazil
Abstract: Technological innovations in vaccinology have recently contributed to bring about novel insights for the vaccine-induced immune response. While the current protocols that use peripheral blood samples may provide abundant data, a range of distinct components of whole blood samples are required and the different anticoagulant systems employed may impair some properties of the biological sample and interfere with functional assays. Although the interference of heparin in functional assays for viral neutralizing antibodies such as the functional plaque-reduction neutralization test (PRNT), considered the gold-standard method to assess and monitor the protective immunity induced by the Yellow fever virus (YFV) vaccine, has been well characterized, the development of pre-analytical treatments is still required for the establishment of optimized protocols. The present study intended to optimize and evaluate the performance of pre-analytical treatment of heparin-collected blood samples with ecteola-cellulose (ECT) to provide accurate measurement of anti-YFV neutralizing antibodies, by PRNT. The study was designed in three steps, including: I. Problem statement; II. Pre-analytical steps; III. Analytical steps. Data confirmed the interference of heparin on PRNT reactivity in a dose-responsive fashion. Distinct sets of conditions for ECT pre-treatment were tested to optimize the heparin removal. The optimized protocol was pre-validated to determine the effectiveness of heparin plasma:ECT treatment to restore the PRNT titers as compared to serum samples. The validation and comparative performance was carried out by using a large range of serum vs heparin plasma:ECT 1:2 paired samples obtained from unvaccinated and 17DD-YFV primary vaccinated subjects. Altogether, the findings support the use of heparin plasma:ECT samples for accurate measurement of anti-YFV neutralizing antibodies.
Keywords: Ecteola-cellulose
Heparin
PRNT
Vaccinology
Yellow fever virus
keywords: Dengue
Issue Date: 2017
Publisher: Elsevier B.V
Citation: AZEVEDO, Ana Carolina Campi et al. Heparin removal by ecteola-cellulose pre-treatment enables the use of plasma samples for accurate measurement of anti-Yellow fever virus neutralizing antibodies. J Immunol Methods., v. 448, p. 9-20, 2017
DOI: 10.1016/j.jim.2017.05.002
ISSN: 0022-1759
Copyright: restricted access
Appears in Collections:Biomanguinhos - Artigos de Periódicos
IRR - Artigos de Periódicos

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