Engineering single-domain antibodies for bioactive paper applications
This thesis is an investigation of the fusion of a carbohydrate-binding module (CBM) to a high-affinity single-domain antibody (sdAb) which binds to bacteriophage M13. This fusion protein was applied to paper filters for detection of M13 phage. A CBM-sdAb fusion protein with nanomolar affinity for immobilized M13 phage was successfully expressed in E. coli. The CBM-sdAb fusion protein was effective in binding M13 phage in water to a cellulose filter paper. However, the sdAb and the CBM-sdAb fusion protein were ineffective in enhancing filter capture of M13 phage particles from water or air, respectively. This research demonstrates that a CBM-sdAb fusion protein will bind simultaneously to cellulose and a model virus. In the future, CBM-sdAb fusion proteins may be useful in the development of ‘bioactive’ paper products capable of detecting and/or inactivating pathogens which are a threat to public health.