A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody

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Standard

A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody. / Pilely, Katrine; Skjoedt, Mikkel-Ole; Nielsen, Christian; Andersen, Thomas Emil; Louise Aabom, Anne; Vitved, Lars; Koch, Claus; Skjødt, Karsten; Palarasah, Yaseelan.

I: Journal of Immunological Methods, Bind 405, 03.2014, s. 87-96.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Pilely, K, Skjoedt, M-O, Nielsen, C, Andersen, TE, Louise Aabom, A, Vitved, L, Koch, C, Skjødt, K & Palarasah, Y 2014, 'A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody', Journal of Immunological Methods, bind 405, s. 87-96. https://doi.org/10.1016/j.jim.2014.01.011

APA

Pilely, K., Skjoedt, M-O., Nielsen, C., Andersen, T. E., Louise Aabom, A., Vitved, L., Koch, C., Skjødt, K., & Palarasah, Y. (2014). A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody. Journal of Immunological Methods, 405, 87-96. https://doi.org/10.1016/j.jim.2014.01.011

Vancouver

Pilely K, Skjoedt M-O, Nielsen C, Andersen TE, Louise Aabom A, Vitved L o.a. A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody. Journal of Immunological Methods. 2014 mar.;405:87-96. https://doi.org/10.1016/j.jim.2014.01.011

Author

Pilely, Katrine ; Skjoedt, Mikkel-Ole ; Nielsen, Christian ; Andersen, Thomas Emil ; Louise Aabom, Anne ; Vitved, Lars ; Koch, Claus ; Skjødt, Karsten ; Palarasah, Yaseelan. / A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody. I: Journal of Immunological Methods. 2014 ; Bind 405. s. 87-96.

Bibtex

@article{61cc4abe2b9746de8d51f64fe59e126e,

title = "A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody",

abstract = "The increasing evidence of the implication of the complement system in the pathogenesis of several diseases has emphasized the need for the development of specific and valid assays, optimized for quantitative detection of complement activation in vivo. In the present study, we have developed a mouse monoclonal antibody (mAb) that is able to detect fluid phase C4c without interference from other products generated from the complement component C4. The C4c specific mAb was tested in different enzyme-linked immunosorbent assay (ELISA) combinations with various types of in vitro activated sera and samples from factor I deficient patients. The specificity of the mAb was further evaluated by immunoprecipitation techniques and by analysis of eluted fragments of C4 after immunoaffinity chromatography. The anti-C4c mAb was confirmed to be C4c specific, as it showed no cross-reactivity with native (un-cleaved) C4, C4b, iC4b, or C4d. Also, no reaction was observed with C4 fragments in factor I deficient plasma or serum samples. We established and validated a sandwich ELISA based on this C4c specific antibody. The normal range of C4c in EDTA/futhan plasma collected from 100 Danish blood donors was measured, with a mean of 0.85mg/L and a range of 0.19-2.21mg/L. We believe that the C4c specific antibody and the ELISA might be important tools in the future assessment of in vivo activation in situations where the classical or the lectin complement pathways are involved in the pathogenesis.",

keywords = "Animals, Antibodies, Monoclonal, Antibody Specificity, Blood Donors, Blotting, Western, Complement Activation, Complement C4, Denmark, Enzyme-Linked Immunosorbent Assay, Humans, Immunoglobulin G, Immunoprecipitation, Mice, Mice, Inbred Strains, Journal Article",

author = "Katrine Pilely and Mikkel-Ole Skjoedt and Christian Nielsen and Andersen, {Thomas Emil} and {Louise Aabom}, Anne and Lars Vitved and Claus Koch and Karsten Skj{\o}dt and Yaseelan Palarasah",

year = "2014",

month = mar,

doi = "10.1016/j.jim.2014.01.011",

language = "English",

volume = "405",

pages = "87--96",

journal = "Journal of Immunological Methods",

issn = "0022-1759",

publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - A specific assay for quantification of human C4c by use of an anti-C4c monoclonal antibody

AU - Pilely, Katrine

AU - Skjoedt, Mikkel-Ole

AU - Nielsen, Christian

AU - Andersen, Thomas Emil

AU - Louise Aabom, Anne

AU - Vitved, Lars

AU - Koch, Claus

AU - Skjødt, Karsten

AU - Palarasah, Yaseelan

PY - 2014/3

Y1 - 2014/3

N2 - The increasing evidence of the implication of the complement system in the pathogenesis of several diseases has emphasized the need for the development of specific and valid assays, optimized for quantitative detection of complement activation in vivo. In the present study, we have developed a mouse monoclonal antibody (mAb) that is able to detect fluid phase C4c without interference from other products generated from the complement component C4. The C4c specific mAb was tested in different enzyme-linked immunosorbent assay (ELISA) combinations with various types of in vitro activated sera and samples from factor I deficient patients. The specificity of the mAb was further evaluated by immunoprecipitation techniques and by analysis of eluted fragments of C4 after immunoaffinity chromatography. The anti-C4c mAb was confirmed to be C4c specific, as it showed no cross-reactivity with native (un-cleaved) C4, C4b, iC4b, or C4d. Also, no reaction was observed with C4 fragments in factor I deficient plasma or serum samples. We established and validated a sandwich ELISA based on this C4c specific antibody. The normal range of C4c in EDTA/futhan plasma collected from 100 Danish blood donors was measured, with a mean of 0.85mg/L and a range of 0.19-2.21mg/L. We believe that the C4c specific antibody and the ELISA might be important tools in the future assessment of in vivo activation in situations where the classical or the lectin complement pathways are involved in the pathogenesis.

AB - The increasing evidence of the implication of the complement system in the pathogenesis of several diseases has emphasized the need for the development of specific and valid assays, optimized for quantitative detection of complement activation in vivo. In the present study, we have developed a mouse monoclonal antibody (mAb) that is able to detect fluid phase C4c without interference from other products generated from the complement component C4. The C4c specific mAb was tested in different enzyme-linked immunosorbent assay (ELISA) combinations with various types of in vitro activated sera and samples from factor I deficient patients. The specificity of the mAb was further evaluated by immunoprecipitation techniques and by analysis of eluted fragments of C4 after immunoaffinity chromatography. The anti-C4c mAb was confirmed to be C4c specific, as it showed no cross-reactivity with native (un-cleaved) C4, C4b, iC4b, or C4d. Also, no reaction was observed with C4 fragments in factor I deficient plasma or serum samples. We established and validated a sandwich ELISA based on this C4c specific antibody. The normal range of C4c in EDTA/futhan plasma collected from 100 Danish blood donors was measured, with a mean of 0.85mg/L and a range of 0.19-2.21mg/L. We believe that the C4c specific antibody and the ELISA might be important tools in the future assessment of in vivo activation in situations where the classical or the lectin complement pathways are involved in the pathogenesis.

KW - Animals

KW - Antibodies, Monoclonal

KW - Antibody Specificity

KW - Blood Donors

KW - Blotting, Western

KW - Complement Activation

KW - Complement C4

KW - Denmark

KW - Enzyme-Linked Immunosorbent Assay

KW - Humans

KW - Immunoglobulin G

KW - Immunoprecipitation

KW - Mice

KW - Mice, Inbred Strains

KW - Journal Article

U2 - 10.1016/j.jim.2014.01.011

DO - 10.1016/j.jim.2014.01.011

M3 - Journal article

C2 - 24472768

VL - 405

SP - 87

EP - 96

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -

ID: 172399187

Niels Bohr Institutet