Binding of complement regulator CFH and CFHL1 by various B.
garinii transformants. B. garinii strains G1, G1/pKFSS1,
G1/pCSPZ, G1/pCSPZ F81A, G1/pCSPZ R139A, G1/pCSPZ Y207A, G1/pCSPZ Y211A,
G1/pCSPZ R139A/Y207A, and G1/pCSPZ Y207A/Y211A incubated in NHS-EDTA were
extensively washed with PBSA containing 0.05% Tween 20, and bound proteins
were eluted using 0.1 m glycine (pH 2.0). Both the last wash
(w) and the eluate (e) fractions obtained from each strain
were separated in a non-reducing conditions 12.5% SDS-PAGE gel, transferred to
nitrocellulose, and probed with either mAb VIG8 specific for SCR 20 of CFH or
αSCR1–4 antiserum specific for the N terminus of CFHL1. As
controls purified CFH and CFHL1 (500 ng each) were loaded on the gel.