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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

TitleDevelopment of an enzyme-linked immunoassay for the quantification of YKL-40 (cartilage gp-39) in guinea pig serum using hen egg yolk antibodies
Authors
Issue Date2001
PublisherElsevier 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?
AbstractAn 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 Identifierhttp://hdl.handle.net/10722/171270
ISSN
2015 Impact Factor: 1.858
2015 SCImago Journal Rankings: 1.053
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorDe Ceuninck, Fen_US
dc.contributor.authorPastoureau, Pen_US
dc.contributor.authorAgnellet, Sen_US
dc.contributor.authorBonnet, Jen_US
dc.contributor.authorVanhoutte, PMen_US
dc.date.accessioned2012-10-30T06:13:04Z-
dc.date.available2012-10-30T06:13:04Z-
dc.date.issued2001en_US
dc.identifier.citationJournal Of Immunological Methods, 2001, v. 252 n. 1-2, p. 153-161en_US
dc.identifier.issn0022-1759en_US
dc.identifier.urihttp://hdl.handle.net/10722/171270-
dc.description.abstractAn 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.languageengen_US
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jimen_US
dc.relation.ispartofJournal of Immunological Methodsen_US
dc.subject.meshAdipokinesen_US
dc.subject.meshAnimalsen_US
dc.subject.meshChickensen_US
dc.subject.meshEgg Yolken_US
dc.subject.meshEnzyme-Linked Immunosorbent Assay - Methods - Standardsen_US
dc.subject.meshGlycoproteins - Blood - Immunologyen_US
dc.subject.meshGuinea Pigsen_US
dc.subject.meshImmunoblotting - Methodsen_US
dc.subject.meshImmunoglobulins - Immunologyen_US
dc.subject.meshLectinsen_US
dc.subject.meshRabbitsen_US
dc.subject.meshRatsen_US
dc.titleDevelopment of an enzyme-linked immunoassay for the quantification of YKL-40 (cartilage gp-39) in guinea pig serum using hen egg yolk antibodiesen_US
dc.typeArticleen_US
dc.identifier.emailVanhoutte, PM:vanhoutt@hku.hken_US
dc.identifier.authorityVanhoutte, PM=rp00238en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/S0022-1759(01)00352-0en_US
dc.identifier.pmid11334975-
dc.identifier.scopuseid_2-s2.0-0035371678en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0035371678&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume252en_US
dc.identifier.issue1-2en_US
dc.identifier.spage153en_US
dc.identifier.epage161en_US
dc.identifier.isiWOS:000168460300016-
dc.publisher.placeNetherlandsen_US
dc.identifier.scopusauthoridDe Ceuninck, F=6602162018en_US
dc.identifier.scopusauthoridPastoureau, P=6603951885en_US
dc.identifier.scopusauthoridAgnellet, S=15723219200en_US
dc.identifier.scopusauthoridBonnet, J=7201770893en_US
dc.identifier.scopusauthoridVanhoutte, PM=7202304247en_US

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