Development of a novel antigen capture-ELISA using IgY against porcine interleukin-6 and its application
Journal of Veterinary Science
; : 337-343, 2004.
Article
em En
| WPRIM
| ID: wpr-79779
Biblioteca responsável:
WPRO
ABSTRACT
Interleukin-6 (IL-6) is introduced as a marker of disease. At present, a variety of method may be used to quantify expression of this protein. Antigen capture-ELISA is a sensitive and accurate quantification method previously used with ovine, rat, and human IL-6 proteins. However, it has never been reported to quantify porcine IL-6 protein using capture ELISA. In this study, we generated and characterized a set of IgY and mono-specific polyclonal antibodies to recombinant porcine IL-6 (rpIL-6), and combining these with a sensitive and specific capture-ELISA for a diagnostic purpose. cDNA encoding the mature protein coding region of porcine IL-6 was cloned and expressed with pQE-30UA expression vector. rpIL-6 was then expressed and purified by using Ni-NTA resin. Protein mass of 24 kDa was found with SDS-PAGE and the identity of the protein was confirmed by Western-blot. Production of polyclonal antibodies against rpIL-6 was performed using the purified rpIL-6 in mice and hens. An antigen capture-ELISA was developed with the antibodies after their extraction. To compare the IL-6 level in the different sanitary state of farms, pig sera were randomly collected and concentration of IL-6 in the sera was measured with the antigen capture-ELISA. The capture-ELISA with the optimal concentration of antibodies, in this study, was able to detect about 10 ng/ml of rpIL-6. IL-6 levels determined with the capture-ELISA in pig sera showed positive correlation with the sanitary states of the farms. These results suggested that the developed antigen capture-ELISA could be a good tool for the screening of microbial infection in pig farms.
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Índice:
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Assunto principal:
Suínos
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Proteínas Recombinantes
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Imunoglobulinas
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Ensaio de Imunoadsorção Enzimática
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Biomarcadores
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Galinhas
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Western Blotting
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Interleucina-6
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Clonagem Molecular
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DNA Complementar
Limite:
Animals
Idioma:
En
Revista:
Journal of Veterinary Science
Ano de publicação:
2004
Tipo de documento:
Article