Our objective is to produces molecular switches that are specifically activated in solution by antibodies and other effectors. Many allosteric enzymes have evolved in nature, but the introduction of a novel regulatory mechanism into a normally unregulated enzyme poses a difficult design problem. We used a combination of rational design, random mutagenesis and screening to engineer an antibody-activated variant of the reporter enzyme beta-glucuronidase (GUS). The activity of the purified GUS variant is increased ~100-fold by the addition of an equimolar concentration of a monoclonal antibody. This molecular switch is modular in design, so it can easily be re-configured for the detection of other peptide-specific antibodies. Such antibody-activated reporters should enable point-of-care serological assays for the detection of infectious diseases.

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