Generation of a C3c specific monoclonal antibody and assessment of C3c as a putative inflammatory marker derived from complement factor C3
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Generation of a C3c specific monoclonal antibody and assessment of C3c as a putative inflammatory marker derived from complement factor C3. / Palarasah, Yaseelan; Skjodt, Karsten; Brandt, Jette; Teisner, Børge; Koch, Claus; Vitved, Lars; Skjoedt, Mikkel-Ole.
I: Journal of Immunological Methods, Bind 362, Nr. 1-2, 31.10.2010, s. 142-50.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Generation of a C3c specific monoclonal antibody and assessment of C3c as a putative inflammatory marker derived from complement factor C3
AU - Palarasah, Yaseelan
AU - Skjodt, Karsten
AU - Brandt, Jette
AU - Teisner, Børge
AU - Koch, Claus
AU - Vitved, Lars
AU - Skjoedt, Mikkel-Ole
N1 - Copyright © 2010 Elsevier B.V. All rights reserved.
PY - 2010/10/31
Y1 - 2010/10/31
N2 - There is a general need for markers of systemic inflammation in acute or chronic diseases, where complement activation is involved. Available methods to monitor complement activation are elaborate and of low sensitivity; they include haemolytic assays (CH50), quantification of fluid phase terminal complex (C5b-C9) and quantification of complement split products by precipitation-in-gel techniques (e.g. C3d). We have developed a mouse monoclonal antibody (mAb) that is able to detect fluid phase C3c without interference from other products generated from the complement component C3. The C3c specific mAb was tested in different ELISA combinations with various types of in vitro activated sera and with plasma or serum samples from factor I deficient patients. The specificity of the mAb was evaluated in immunoprecipitation techniques and by analysis of eluted fragments of C3 after immunoaffinity chromatography. The C3c mAb was confirmed to be C3c specific, as it showed no cross-reactivity with native (un-cleaved) C3, with C3b, iC3b, or with C3d. Also, no significant reaction was observed with C3 fragments in factor I deficient sera or plasma. This antibody forms the basis for the generation of a robust ELISA that allows for a quick and reliable evaluation of complement activation and consumption as a marker for inflammatory processes. We established the C3c plasma range in 100 healthy Danish blood donors with a mean of 3.47 μg/ml and a range of 2.12-4.92 μg/ml. We believe that such an antibody might be of potential value in the assessment of in vivo complement activity during the inflammatory processes.
AB - There is a general need for markers of systemic inflammation in acute or chronic diseases, where complement activation is involved. Available methods to monitor complement activation are elaborate and of low sensitivity; they include haemolytic assays (CH50), quantification of fluid phase terminal complex (C5b-C9) and quantification of complement split products by precipitation-in-gel techniques (e.g. C3d). We have developed a mouse monoclonal antibody (mAb) that is able to detect fluid phase C3c without interference from other products generated from the complement component C3. The C3c specific mAb was tested in different ELISA combinations with various types of in vitro activated sera and with plasma or serum samples from factor I deficient patients. The specificity of the mAb was evaluated in immunoprecipitation techniques and by analysis of eluted fragments of C3 after immunoaffinity chromatography. The C3c mAb was confirmed to be C3c specific, as it showed no cross-reactivity with native (un-cleaved) C3, with C3b, iC3b, or with C3d. Also, no significant reaction was observed with C3 fragments in factor I deficient sera or plasma. This antibody forms the basis for the generation of a robust ELISA that allows for a quick and reliable evaluation of complement activation and consumption as a marker for inflammatory processes. We established the C3c plasma range in 100 healthy Danish blood donors with a mean of 3.47 μg/ml and a range of 2.12-4.92 μg/ml. We believe that such an antibody might be of potential value in the assessment of in vivo complement activity during the inflammatory processes.
KW - Animals
KW - Antibodies, Monoclonal
KW - Antibody Specificity
KW - Biomarkers
KW - Blood Donors
KW - Complement Activation
KW - Complement C3c
KW - Denmark
KW - Enzyme-Linked Immunosorbent Assay
KW - Female
KW - Humans
KW - Inflammation
KW - Inflammation Mediators
KW - Male
KW - Mice
KW - Mice, Inbred BALB C
KW - Sensitivity and Specificity
KW - Journal Article
U2 - 10.1016/j.jim.2010.09.024
DO - 10.1016/j.jim.2010.09.024
M3 - Journal article
C2 - 20869965
VL - 362
SP - 142
EP - 150
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -
ID: 172399698