TY - JOUR
T1 - Anti-peptide antibodies
T2 - A tool for detecting IL-8 in salmonids
AU - Santana, Paula
AU - Palacios, Claudia
AU - Narváez, Edgar
AU - Guzmán, Fanny
AU - Gallardo, José A.
AU - Mercado, Luis
PY - 2012/9/15
Y1 - 2012/9/15
N2 - Background: Interleukin 8 is a chemokine that is produced by several types of cells, like macrophages and has chemotactic activity in particular on neutrophils, playing a key role during the inflammatory process. It has been demonstrated at the molecular level that this molecule is present and conserved in several vertebrate groups, pointing its importance. Analysis of the amino acid sequence of IL-8, projected from cDNA of Salmo salar, presents homology with the sequences of mammals, poultry and lamprey, indicating the presence of a homologous molecule in higher fish. However, there is no information at protein level, which allows characterizing the regulatory role of this molecule during the immune response in fish. Results: In this work, we designed and synthesized an epitope peptide of 10 residues with a purity of 95% and mass of 1158.7 kDa, which showed a random coil structure. From this peptide it was able to generate a polyclonal mono-specific antibody which was capable of detecting the whole molecule of IL-8 in tissue and cellular model of salmonids. Conclusions: The resulting antibody is a versatile tool for detecting IL-8 by different immune techniques such as ELISA, dot blot, western blotting and immunocytofluorescence. Analysis of IL-8 at proteomic level is a useful method for characterizing immune properties of this molecule in fish.
AB - Background: Interleukin 8 is a chemokine that is produced by several types of cells, like macrophages and has chemotactic activity in particular on neutrophils, playing a key role during the inflammatory process. It has been demonstrated at the molecular level that this molecule is present and conserved in several vertebrate groups, pointing its importance. Analysis of the amino acid sequence of IL-8, projected from cDNA of Salmo salar, presents homology with the sequences of mammals, poultry and lamprey, indicating the presence of a homologous molecule in higher fish. However, there is no information at protein level, which allows characterizing the regulatory role of this molecule during the immune response in fish. Results: In this work, we designed and synthesized an epitope peptide of 10 residues with a purity of 95% and mass of 1158.7 kDa, which showed a random coil structure. From this peptide it was able to generate a polyclonal mono-specific antibody which was capable of detecting the whole molecule of IL-8 in tissue and cellular model of salmonids. Conclusions: The resulting antibody is a versatile tool for detecting IL-8 by different immune techniques such as ELISA, dot blot, western blotting and immunocytofluorescence. Analysis of IL-8 at proteomic level is a useful method for characterizing immune properties of this molecule in fish.
KW - Anti-IL-8
KW - Fish antibodies
KW - Fish chemokine
KW - Rainbow trout
KW - Synthetic peptide epitope
UR - http://www.scopus.com/inward/record.url?scp=84866613627&partnerID=8YFLogxK
U2 - 10.2225/vol15-issue5-fulltext-15
DO - 10.2225/vol15-issue5-fulltext-15
M3 - Article
AN - SCOPUS:84866613627
SN - 0717-3458
VL - 15
SP - 20
JO - Electronic Journal of Biotechnology
JF - Electronic Journal of Biotechnology
IS - 5
ER -