Publicación:
Single-chain antibodies from alpaca for the detection of Fasciola hepatica antigens [Anticuerpos de cadena única de alpaca para la detección de antígenos de Fasciola hepatica]
Single-chain antibodies from alpaca for the detection of Fasciola hepatica antigens [Anticuerpos de cadena única de alpaca para la detección de antígenos de Fasciola hepatica]
| dc.contributor.author | Barreto T. | es_PE |
| dc.contributor.author | Alfonso Y. | es_PE |
| dc.contributor.author | Lafaye P. | es_PE |
| dc.contributor.author | García Lazaro M.P. | es_PE |
| dc.contributor.author | Perez L.A. | es_PE |
| dc.contributor.author | Herrera-Velit P. | es_PE |
| dc.contributor.author | Espinoza J.R. | es_PE |
| dc.date.accessioned | 2024-05-30T23:13:38Z | |
| dc.date.available | 2024-05-30T23:13:38Z | |
| dc.date.issued | 2018 | |
| dc.description | Dr. J. Rodriguez, E. Gushiken por el modelamiento del VHH-ES1. El vector pHEN2 fue donado por el Dr. D. Baty y Dr. D. Kastelic (Protein Biotechnology Group, Babraham Bioscience Technologies, UK). TB realizó una estadía en el Instituto Pasteur, Francia bajo la supervisión del Dr. P. Lafaye. Fuentes de financiamiento: El trabajo fue financiado por el Programa Nacional de Innovación para la Competitividad y Productividad (Innóvate Perú), contrato 123-FINCyT-IA-2013, «Generación de anticuerpos de dominio-único de alpacas para el diagnóstico y tratamiento de enfermedades hepáticas». YA recibió financiamiento por el proyecto Postdoctoral FONDECYT «Generación de Anticuerpos de dominio único de alpacas para el diagnóstico y tratamiento de enfermedades hepáticas», EF015-2013. | |
| dc.description.abstract | Objectives. To produce recombinant single-chain antibodies from alpaca that will bind to the excreted-secreted (ES) Fasciola hepatica antigen with high affinity and specificity, so as to develop new diagnostic technologies of human and animal fascioliasis. Materials and Methods. A gene bank of DNA of the variable dominions of heavy single-chain antibodies (VHH) has been created, based on mononuclear cells of peripheral blood of an alpaca immunized with the ES antigen of F. hepatica. The gene bank was screened with the ES antigen by differential phage display, selecting ten VHH that bind specifically to ES. The anti-ES VHH was cloned in an expression vector, the recombinant protein (VHH-ES1) of 15.3 kDa was produced by fermentation in E. coli and purified to homogeneity by affinity chromatography. The binding of VHH-ES1 to the ES antigen was evaluated by ELISA using VHH-ES1 as capture antibody, policlonal anti-ES serum of rabbit and conjugated rabbit anti IgG with radish peroxidase. Results. A VHH that binds to the ES antigen (VHH-ES1) has been identified through differential phage display and produced by fermentation in E. coli; this corresponds to an antibody of the long-hinge IgG2 subclass. The binding of the VHH-ES1 antibody to the antigen shows linearity with respect to the concentration of ES in the 50-5,000 ng/mL range and the limit of detection value of the antigen is in the 30-170 ng/mL range of ES (R2=0.99). Conclusions. The VHH-ES1 binds with affinity and specificity to the ES antigen of F. hepatica and is a promissory antibody to be assessed for the development of new fascioliasis diagnostic technologies. © 2018, Instituto Nacional de Salud. All rights reserved. | |
| dc.description.sponsorship | Consejo Nacional de Ciencia, Tecnología e Innovación Tecnológica - Concytec | |
| dc.identifier.doi | https://doi.org/10.17843/rpmesp.2018.354.3101 | |
| dc.identifier.scopus | 2-s2.0-85061121219 | |
| dc.identifier.uri | https://hdl.handle.net/20.500.12390/2306 | |
| dc.language.iso | eng | |
| dc.publisher | Instituto Nacional de Salud | |
| dc.relation.ispartof | Revista Peruana de Medicina Experimental y Salud Publica | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/4.0/ | |
| dc.subject | Single-domain antibodies | |
| dc.subject | Antibodies | es_PE |
| dc.subject | Camelids | es_PE |
| dc.subject | Fascioliasis | es_PE |
| dc.subject | New World | es_PE |
| dc.subject | Peru (source MeSH NLM) | es_PE |
| dc.subject.ocde | http://purl.org/pe-repo/ocde/ford#3.04.03 | |
| dc.title | Single-chain antibodies from alpaca for the detection of Fasciola hepatica antigens [Anticuerpos de cadena única de alpaca para la detección de antígenos de Fasciola hepatica] | |
| dc.type | info:eu-repo/semantics/article | |
| dspace.entity.type | Publication |