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- Publisher Website: 10.1016/S0022-1759(01)00352-0
- Scopus: eid_2-s2.0-0035371678
- PMID: 11334975
- WOS: WOS:000168460300016
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Article: Development of an enzyme-linked immunoassay for the quantification of YKL-40 (cartilage gp-39) in guinea pig serum using hen egg yolk antibodies
Title | Development of an enzyme-linked immunoassay for the quantification of YKL-40 (cartilage gp-39) in guinea pig serum using hen egg yolk antibodies |
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Authors | |
Keywords | Chitinase Disease marker Egg yolk antibodies ELISA Guinea pig Osteoarthritis YKL-40/HC gp-39 |
Issue Date | 2001 |
Publisher | Elsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jim |
Citation | Journal Of Immunological Methods, 2001, v. 252 n. 1-2, p. 153-161 How to Cite? |
Abstract | An indirect competition immunoassay for the quantification of YKL-40 (cartilage gp-39, Chondrex) in guinea pig serum has been developed using egg yolk antibodies (IgY). The immune response of hens to YKL-40 was verified by immunoblot analyses. Highly specific antibodies were obtained 30 days after the first injection. The ELISA was developed in 96-well microtiter plates with quadruplicate determinations for each point. The assay was based on the ability of YKL-40 present in serum to displace the binding of antibodies to the coated antigen. An inhibition mixture containing standard YKL-40 or guinea pig serum, diluted 1/5, and primary antibodies, diluted 1/5000, was allowed to equilibrate for 2 h at room temperature and dispensed for 16 h at 4°C in wells coated with 1 μg/ml of YKL-40. Detection was achieved by the addition of rabbit anti-chicken antibodies conjugated to peroxidase followed by tetramethylbenzidine. Specificity was assessed by parallelism between a dilution curve of serum and standard YKL-40. The sensitivity of detection was 10 ng/ml. Intra- and interassay coefficients of variation were both 8.7%. The analytical recovery was 101.5 ± 5.4% (mean ± standard deviation (SD), n = 9). The YKL-40 concentration in serum from 12 adult guinea pigs was 330 ± 216 ng/ml (mean ± SD) with a lower value of 164 ng/ml and an upper value of 982 ng/ml. In contrast to the rat, a dilution curve of rabbit serum gave parallelism with the guinea pig standard, suggesting recognition of a similar epitope. Possible applications of the assay in the guinea pig include disease models where YKL-40 is overexpressed and could be used as a marker, i.e. osteoarthritis, rheumatoid arthritis, cancer, liver fibrosis, atherosclerosis and more generally, pathologies with increased tissue remodeling. © 2001 Elsevier Science B.V. |
Persistent Identifier | http://hdl.handle.net/10722/171270 |
ISSN | 2023 Impact Factor: 1.6 2023 SCImago Journal Rankings: 0.555 |
ISI Accession Number ID | |
References |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | De Ceuninck, F | en_US |
dc.contributor.author | Pastoureau, P | en_US |
dc.contributor.author | Agnellet, S | en_US |
dc.contributor.author | Bonnet, J | en_US |
dc.contributor.author | Vanhoutte, PM | en_US |
dc.date.accessioned | 2012-10-30T06:13:04Z | - |
dc.date.available | 2012-10-30T06:13:04Z | - |
dc.date.issued | 2001 | en_US |
dc.identifier.citation | Journal Of Immunological Methods, 2001, v. 252 n. 1-2, p. 153-161 | en_US |
dc.identifier.issn | 0022-1759 | en_US |
dc.identifier.uri | http://hdl.handle.net/10722/171270 | - |
dc.description.abstract | An indirect competition immunoassay for the quantification of YKL-40 (cartilage gp-39, Chondrex) in guinea pig serum has been developed using egg yolk antibodies (IgY). The immune response of hens to YKL-40 was verified by immunoblot analyses. Highly specific antibodies were obtained 30 days after the first injection. The ELISA was developed in 96-well microtiter plates with quadruplicate determinations for each point. The assay was based on the ability of YKL-40 present in serum to displace the binding of antibodies to the coated antigen. An inhibition mixture containing standard YKL-40 or guinea pig serum, diluted 1/5, and primary antibodies, diluted 1/5000, was allowed to equilibrate for 2 h at room temperature and dispensed for 16 h at 4°C in wells coated with 1 μg/ml of YKL-40. Detection was achieved by the addition of rabbit anti-chicken antibodies conjugated to peroxidase followed by tetramethylbenzidine. Specificity was assessed by parallelism between a dilution curve of serum and standard YKL-40. The sensitivity of detection was 10 ng/ml. Intra- and interassay coefficients of variation were both 8.7%. The analytical recovery was 101.5 ± 5.4% (mean ± standard deviation (SD), n = 9). The YKL-40 concentration in serum from 12 adult guinea pigs was 330 ± 216 ng/ml (mean ± SD) with a lower value of 164 ng/ml and an upper value of 982 ng/ml. In contrast to the rat, a dilution curve of rabbit serum gave parallelism with the guinea pig standard, suggesting recognition of a similar epitope. Possible applications of the assay in the guinea pig include disease models where YKL-40 is overexpressed and could be used as a marker, i.e. osteoarthritis, rheumatoid arthritis, cancer, liver fibrosis, atherosclerosis and more generally, pathologies with increased tissue remodeling. © 2001 Elsevier Science B.V. | en_US |
dc.language | eng | en_US |
dc.publisher | Elsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jim | en_US |
dc.relation.ispartof | Journal of Immunological Methods | en_US |
dc.subject | Chitinase | - |
dc.subject | Disease marker | - |
dc.subject | Egg yolk antibodies | - |
dc.subject | ELISA | - |
dc.subject | Guinea pig | - |
dc.subject | Osteoarthritis | - |
dc.subject | YKL-40/HC gp-39 | - |
dc.subject.mesh | Adipokines | en_US |
dc.subject.mesh | Animals | en_US |
dc.subject.mesh | Chickens | en_US |
dc.subject.mesh | Egg Yolk | en_US |
dc.subject.mesh | Enzyme-Linked Immunosorbent Assay - Methods - Standards | en_US |
dc.subject.mesh | Glycoproteins - Blood - Immunology | en_US |
dc.subject.mesh | Guinea Pigs | en_US |
dc.subject.mesh | Immunoblotting - Methods | en_US |
dc.subject.mesh | Immunoglobulins - Immunology | en_US |
dc.subject.mesh | Lectins | en_US |
dc.subject.mesh | Rabbits | en_US |
dc.subject.mesh | Rats | en_US |
dc.title | Development of an enzyme-linked immunoassay for the quantification of YKL-40 (cartilage gp-39) in guinea pig serum using hen egg yolk antibodies | en_US |
dc.type | Article | en_US |
dc.identifier.email | Vanhoutte, PM:vanhoutt@hku.hk | en_US |
dc.identifier.authority | Vanhoutte, PM=rp00238 | en_US |
dc.description.nature | link_to_subscribed_fulltext | en_US |
dc.identifier.doi | 10.1016/S0022-1759(01)00352-0 | en_US |
dc.identifier.pmid | 11334975 | - |
dc.identifier.scopus | eid_2-s2.0-0035371678 | en_US |
dc.relation.references | http://www.scopus.com/mlt/select.url?eid=2-s2.0-0035371678&selection=ref&src=s&origin=recordpage | en_US |
dc.identifier.volume | 252 | en_US |
dc.identifier.issue | 1-2 | en_US |
dc.identifier.spage | 153 | en_US |
dc.identifier.epage | 161 | en_US |
dc.identifier.isi | WOS:000168460300016 | - |
dc.publisher.place | Netherlands | en_US |
dc.identifier.scopusauthorid | De Ceuninck, F=6602162018 | en_US |
dc.identifier.scopusauthorid | Pastoureau, P=6603951885 | en_US |
dc.identifier.scopusauthorid | Agnellet, S=15723219200 | en_US |
dc.identifier.scopusauthorid | Bonnet, J=7201770893 | en_US |
dc.identifier.scopusauthorid | Vanhoutte, PM=7202304247 | en_US |
dc.identifier.issnl | 0022-1759 | - |