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Monoclonal antibody-mediated inhibition of complement to prevent pathology

Monday 18 August 2014

Dr Peter Boross, Prof Dr Erik Hack

Excessive complement activation underlies several pathologies, such as atypical haemolytic uremic syndrome (aHUS), antibody-mediated rejection (AMR) during kidney transplantation or haemolytic anaemia. For instance, in haemolytic anaemia, autoantibodies directed against erythrocytes cause pathology via either direct complement-mediated lysis (intravascular haemolysis) or phagocytosis of opsonized erythrocytes by macrophages (extravascular haemolysis). Inhibition of complement activation is therefore an attractive method for the treatment of these conditions, as shown by the successful introduction of eculizumab, an anti-C5 monoclonal antibody (mAb) to use in aHUS.
We have developed a novel therapeutic mAb that effectively inhibits complement activation via the classical pathway in several in vitro assays. This mAb also effectively inhibits AMR in vitro. We will set up and optimize in vitro assays with primary erythrocytes and human monocytes / macrophages to assess the efficacy of this novel mAb in inhibiting intra- and extravascular haemolysis.
In addition, we have developed a panel of novel mAbs to inhibit the alternative pathway of complement. These mAbs will be characterized using various biochemical and ELISA-based complement activation assays. The epitopes of these novel mAbs will be mapped using existing domain-swap mutants and/or newly generated point mutants. In addition, the generation of novel complement-inhibitory mAbs will also be considered.
Using these novel tools and existing mAbs that target complement components we aim to unravel the complex pathways that underlie complement-induced pathology.

Cell-culture, isolation of monocytes from peripheral blood, FACS, ELISA, microscopy, antibody purification, SDS-PAGE, Western blot, molecular cloning

6-9 month (preferably 9 months)

Dr. Peter Boross, p.boross@umcutrecht.nl
Prof Dr Erik Hack, c.e.hack@umcutrecht.nl
Dr. Kristin Denzer, k.denzer@umcutrecht.nl, 088 75 643 68