Novel monoclonal antibodies against mouse C3 interfering with complement activation: Description of fine specificity and applications to various immunoassays

Dimitrios Mastellos, J. Prechl, Glória László, Krisztián Papp, Eszter Oláh, Emelia Argyropoulos, Silvia Franchini, Ruxandra Tudoran, Maciej Markiewski, John D. Lambris, A. Erdei

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

The role of complement proteins in various pathophysiological settings has been studied primarily using mouse models of disease. However, the specific contribution of C3-derived fragments to these biologic processes has not been addressed in a rigorous manner because of a lack of antibodies that can selectively recognize mouse C3 or any of its degradation fragments. Here we report the generation and characterization of a panel of rat monoclonal antibodies reacting with mouse C3 and its degradation products. We describe their performance in various immunological assays such as ELISA, Western blotting, flow cytometry and immunohistochemistry. Of all the antibodies generated, one selectively recognized the C3a anaphylatoxin, and all other reacted with C3c. Furthermore, two monoclonal antibodies preferentially reacted with the cleaved C3 fragments C3b/iC3b/C3c but not native C3. Except for the one recognizing C3a, all antibodies were suitable for detecting C3 deposited on cells and tissues, two effectively inhibited the hemolytic activity of mouse complement and one enhanced C3-deposition to the cell membrane. These novel monoclonal antibodies may serve as useful reagents for elucidating functions mediated by C3-derived fragments in various pathophysiological conditions.

Original languageEnglish
Pages (from-to)1213-1221
Number of pages9
JournalMolecular Immunology
Volume40
Issue number16
DOIs
Publication statusPublished - Mar 2004

Fingerprint

Complement Activation
Immunoassay
Monoclonal Antibodies
Antibodies
Anaphylatoxins
Complement C3b
Flow Cytometry
Complement System Proteins
Western Blotting
Enzyme-Linked Immunosorbent Assay
Immunohistochemistry
Cell Membrane

Keywords

  • C3
  • ELISA
  • Monoclonal antibody
  • Mouse

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

Novel monoclonal antibodies against mouse C3 interfering with complement activation : Description of fine specificity and applications to various immunoassays. / Mastellos, Dimitrios; Prechl, J.; László, Glória; Papp, Krisztián; Oláh, Eszter; Argyropoulos, Emelia; Franchini, Silvia; Tudoran, Ruxandra; Markiewski, Maciej; Lambris, John D.; Erdei, A.

In: Molecular Immunology, Vol. 40, No. 16, 03.2004, p. 1213-1221.

Research output: Contribution to journalArticle

Mastellos, Dimitrios ; Prechl, J. ; László, Glória ; Papp, Krisztián ; Oláh, Eszter ; Argyropoulos, Emelia ; Franchini, Silvia ; Tudoran, Ruxandra ; Markiewski, Maciej ; Lambris, John D. ; Erdei, A. / Novel monoclonal antibodies against mouse C3 interfering with complement activation : Description of fine specificity and applications to various immunoassays. In: Molecular Immunology. 2004 ; Vol. 40, No. 16. pp. 1213-1221.
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