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| Natura: | Artículo científico |
| Lingua: | es |
| Pubblicazione: |
Universidad Austral de Chile
2015
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| Accesso online: | https://www.redalyc.org/articulo.oa?id=173042665008 |
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Sommario:
- Identificación de nuevos péptidos de una biblioteca de despliegue en fagos con un anticuerpo monoclonal específico a la proteína N del virus PRRS A Villa-Mancera A Reynoso-Paloma F Utrera-Quintana A Córdova-Izquierdo J Olivares-Pérez M Zambrano-González A Trejo-Córdova L Carreón-Luna Veterinaria PRRS N protein mimotopes phage display The aim of the present study was to select phage clones from a combinatorial library of filamentous phage that mimic the N protein of Porcine Reproductive and Respiratory Syndrome Virus (PRRS) using a monoclonal antibody (SDOW17). An combinatorial library of random peptide 7-mer cysteine-constrained fused to a minor coat protein (pIII) of the M13 phage was used. To determine the effect of enrichment after each round of selection, the eluted phage were titrated, bacteriophages increased from 4,8 x 10 3 plaque-forming units (pfu) in the first round to 3,2 x 10 5 pfu in the third round. After three rounds of panning 32 phage clones were randomly picked, each individual clone was amplified, purified and titred. The binding reactivity of phage clones with anti-PRRS antibodies was determined using a phage ELISA. Alignments of the selected clones with the published sequences of the N protein of PRRS virus were located at the amino terminal region and middle portion of the protein. The consensus amino acid residues of the 32 mimotopes sequenced was NYRYQ. Mimotope sequences were used to calculate the epitopes of the nucleocapsid protein by the web tool of Peptiope server with PepSurf algorithm. Two conformational epitopes against monoclonal antibody anti-protein N were identified, located in a different position and appeared mainly as an α -helix. The selected mimotopes and conformational epitopes could be regarded as informative for diagnosis of disease. 2015 artículo científico 0301-732X https://www.redalyc.org/articulo.oa?id=173042665008 es http://www.redalyc.org/revista.oa?id=1730 Archivos de Medicina Veterinaria application/pdf Universidad Austral de Chile Archivos de Medicina Veterinaria (Chile) Num.3 Vol.47