Development of a novel antigen capture-ELISA using IgY against porcine interleukin-6 and its application.

Deog Yong Lee, Young Wook Cho, Sang Gyun Kang, SungJae Shin, Han Sang Yoo

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)337-343
Number of pages7
JournalJournal of veterinary science (Suwon-si, Korea)
Volume5
Issue number4
Publication statusPublished - 2004 Jan 1

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interleukin-6
Interleukin-6
Swine
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
antigens
Antigens
swine
Proteins
polyclonal antibodies
Antibodies
farms
IgY
Serum
proteins
antibodies
Open Reading Frames
Antibody Formation
Polyacrylamide Gel Electrophoresis
Sheep

All Science Journal Classification (ASJC) codes

  • veterinary(all)

Cite this

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title = "Development of a novel antigen capture-ELISA using IgY against porcine interleukin-6 and its application.",
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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Development of a novel antigen capture-ELISA using IgY against porcine interleukin-6 and its application. / Lee, Deog Yong; Cho, Young Wook; Kang, Sang Gyun; Shin, SungJae; Yoo, Han Sang.

In: Journal of veterinary science (Suwon-si, Korea), Vol. 5, No. 4, 01.01.2004, p. 337-343.

Research output: Contribution to journalArticle

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