A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5 '-diphosphocholine coupled dendrimers

Peter M. H. Heegaard, H. G. Pedersen, A. L. Jensen, Ulrik Boas

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP. The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity.
    Original languageEnglish
    JournalJournal of Immunological Methods
    Volume343
    Issue number2
    Pages (from-to)112-118
    ISSN0022-1759
    DOIs
    Publication statusPublished - 2009

    Keywords

    • Dendrimer
    • Acute phase protein
    • Pig CRP
    • Phosphocholine
    • Human CRP
    • Immunoassay

    Cite this

    @article{757d2ddb54a84404b9765c8b4bf5aba5,
    title = "A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5 '-diphosphocholine coupled dendrimers",
    abstract = "C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP. The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity.",
    keywords = "Dendrimer, Acute phase protein, Pig CRP, Phosphocholine, Human CRP, Immunoassay",
    author = "Heegaard, {Peter M. H.} and Pedersen, {H. G.} and Jensen, {A. L.} and Ulrik Boas",
    year = "2009",
    doi = "10.1016/j.jim.2009.02.002",
    language = "English",
    volume = "343",
    pages = "112--118",
    journal = "Journal of Immunological Methods",
    issn = "0022-1759",
    publisher = "Elsevier",
    number = "2",

    }

    A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5 '-diphosphocholine coupled dendrimers. / Heegaard, Peter M. H.; Pedersen, H. G.; Jensen, A. L.; Boas, Ulrik.

    In: Journal of Immunological Methods, Vol. 343, No. 2, 2009, p. 112-118.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5 '-diphosphocholine coupled dendrimers

    AU - Heegaard, Peter M. H.

    AU - Pedersen, H. G.

    AU - Jensen, A. L.

    AU - Boas, Ulrik

    PY - 2009

    Y1 - 2009

    N2 - C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP. The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity.

    AB - C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP. The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity.

    KW - Dendrimer

    KW - Acute phase protein

    KW - Pig CRP

    KW - Phosphocholine

    KW - Human CRP

    KW - Immunoassay

    U2 - 10.1016/j.jim.2009.02.002

    DO - 10.1016/j.jim.2009.02.002

    M3 - Journal article

    C2 - 19236874

    VL - 343

    SP - 112

    EP - 118

    JO - Journal of Immunological Methods

    JF - Journal of Immunological Methods

    SN - 0022-1759

    IS - 2

    ER -