Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/13829
Title: Novel Camelid Antibody Fragments Targeting Recombinant Nucleoprotein of Araucaria hantavirus: A Prototype for an Early Diagnosis of Hantavirus Pulmonary Syndrome
Authors: Pereira, Soraya S.
Dill, Leandro S. Moreira
Morais, Michelle S. S.
Prado, Nidiane D. R.
Barros, Marcos L.
Koishi, Andrea C.
Mazarrotto, Giovanny A. C. A.
Gonçalves, Giselle M.
Zuliani, Juliana P.
Calderon, Leonardo A.
Soares, Andreimar M.
Silva, Luiz H. Pereira da
Santos, Claudia N. Duarte dos
Fernandes, Carla F. C.
Stabeli, Rodrigo G.
Affilliation: Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PA, Brazil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PA, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil. / Universidade Federal de Rondônia. Departamento de Medicina. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil. / Universidade Federal de Rondônia. Departamento de Medicina. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PA, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil. / Centro de Pesquisa em Medicina Tropical. Porto Velho, RO, Brazil.
Fundação Oswaldo Cruz. Porto Velho, RO, Brazil. / Universidade Federal de Rondônia. Departamento de Medicina. Porto Velho, RO, Brazil.
Abstract: In addition to conventional antibodies, camelids produce immunoglobulins G composed exclusively of heavy chains in which the antigen binding site is formed only by single domains called VHH. Their particular characteristics make VHHs interesting tools for drug-delivery, passive immunotherapy and high-throughput diagnosis. Hantaviruses are rodent-borne viruses of the Bunyaviridae family. Two clinical forms of the infection are known. Hemorrhagic Fever with Renal Syndrome (HFRS) is present in the Old World, while Hantavirus Pulmonary Syndrome (HPS) is found on the American continent. There is no specific treatment for HPS and its diagnosis is carried out by molecular or serological techniques, using mainly monoclonal antibodies or hantavirus nucleoprotein (N) to detect IgM and IgG in patient serum. This study proposes the use of camelid VHHs to develop alternative methods for diagnosing and confirming HPS. Phage display technology was employed to obtain VHHs. After immunizing one Lama glama against the recombinant N protein (prND85) of a Brazilian hantavirus strain, VHH regions were isolated to construct an immune library. VHHs were displayed fused to the M13KO7 phage coat protein III and the selection steps were performed on immobilized prND85. After selection, eighty clones recognized specifically the N protein. These were sequenced, grouped based mainly on the CDRs, and five clones were analyzed by western blot (WB), surface plasmon resonance (SPR) device, and ELISA. Besides the ability to recognize prND85 by WB, all selected clones showed affinity constants in the nanomolar range. Additionaly, the clone KC329705 is able to detect prND85 in solution, as well as the native viral antigen. Findings support the hypothesis that selected VHHs could be a powerful tool in the development of rapid and accurate HPS diagnostic assays, which are essential to provide supportive care to patients and reduce the high mortality rate associated with hantavirus infections.
Keywords: Camelid
Nucleoprotein
Hantavirus
Pulmonary Syndrome
Issue Date: 2014
Publisher: Plos One
Citation: SOARES, A. M. et al. Novel Camelid Antibody Fragments Targeting Recombinant Nucleoprotein of Araucaria hantavirus: A Prototype for an Early Diagnosis of Hantavirus Pulmonary Syndrome. Plos One, v. 9, p. e108067, 2014.
DOI: 10.1371/journal.pone.0108067
Copyright: open access
Appears in Collections:PR - ICC - Artigos de Periódicos
RO - Artigos de Periódicos

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