title: Antibody-mediates inhibition of human C1s and the classical complement pathway.
abstract: Disregulation of complement activation plays a critical role in numerous inflammatory diseases and therefore, inhibition of the complement pathway is of great therapeutic interest. In the classical complement pathway, immune complexes formed by IgM, IgG1, IgG2 and IgG3 antibodies result in the activation of the C1s protease that in turn cleaves C4 and then C4-bound-C2 yielding the proteolytic fragments C4b and C2a which associate to form a C3 convertase enzyme. We report here the engineering of a potent human antibody inhibitor of C1s protease activity. Phage panning of a very large synthetic (F(AB)) antibody fragment library using a truncated version of C1s, comprising the second CCP domain and serine protease domain (CCP₂-SP) and expressed in insect cells, resulted in the isolation of a F(AB) that inhibited the catalytic activity of C1s. An affinity matured variant of the F(AB) format antibody displaying subnanomolar K(D) for C1s was shown to exhibit >80% inhibition of C2 processing at a 5:1 antibody:C1s molar ratio. We show that this engineered antibody, D.35, displays potent inhibition of complement deposition and lysis of Ramos cells by the anti-CD20 therapeutic antibody rituximab relative to the approved, but less-specific, human plasma-derived C1-inhibitor (CINRYZE). C1s inhibitory antibodies should be useful for delineating the role of the classical pathway in disease models and may hold promise as therapeutic agents.
authors: Department of Chemical Engineering, University of Texas at Austin, Austin, TX, USA.
authors: Sean
authors: S
authors: Carroll
authors: George
authors: G
authors: Georgiou
mesh_terms: D000906:Antibodies / Q000276:immunology*; D003167:Complement Activation / Q000276:immunology*; D003173:Complement C1s / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D003181:Complement C4 / Q000276:immunology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*
chemical_list: D000906:Antibodies; D003181:Complement C4; D007140:Immunoglobulin Fab Fragments; D003173:Complement C1s