imm.13228.pdf (3.65 MB)
Characterising the original anti-C5 function-blocking antibody, BB5.1, for species specificity, mode of action and interactions with C5.
journal contributionposted on 2020-10-07, 13:07 authored by Wioleta M Zelek, Georgina E Menzies, Andrea Brancale, Brigitta Stockinger, Bryan Paul Morgan
The implication of complement in multiple diseases over the last twenty years has fuelled interest in developing anti-complement drugs. To date, the focus has been on C5; blocking cleavage of C5 prevents formation of two pro-inflammatory activities, C5a anaphylatoxin and membrane attack complex. The concept of C5 blockade to inhibit inflammation dates back thirty years to the description of BB5.1, an anti-C5 blocking monoclonal antibody raised in C5-deficient mice. This antibody proved an invaluable tool to demonstrate complement involvement in mouse disease models and catalysed enthusiasm for anti-complement drug development, culminating in the anti-human C5 monoclonal antibody ecuizumab, the most successful anti-complement drug to date, already in the clinic for several rare diseases. Despite its key role in providing proof-of-concept for C5 blockade, the mechanism of BB5.1 inhibition remains poorly understood. Here we characterised BB5.1 cross-species inhibition, C5 binding affinity and chain specificity. BB5.1 efficiently inhibited C5 in mouse serum but not in human or other rodent sera; it prevented C5 cleavage and C5a generation. BB5.1 bound the C5 α-chain with high affinity and slow off-rate. BB5.1 complementarity determining regions were obtained and docking algorithms used to predict the likely binding interface on mouse C5.