Abstract

Gram-positive bacterial pathogens express immunoglobulin (Ig) binding proteins that perturb Fc-dependent functions such as the interaction with complement or phagocytic Fc-receptors. The possession of such molecules by gram-negative bacteria, including Escherichia coli ( E. coli), has also been documented. In many such studies, the detection of Ig binding has relied on the use of polyclonal antibodies as detecting reagents. These are not ideal since such preparations may be contaminated with E. coli specific IgG, allowing for the potential misinterpretation of specific F(ab′) 2 binding as non-specific Fc mediated binding. Here we use mono-specific recombinant antibodies to develop a novel assay for Ig binding non reliant on traditional polyclonal antibodies, allowing us to demonstrate the unequivocal absence of Fc binding proteins from E. coli.

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