Phages as a tool in receptor development for biosensing
Claudia Pahlke, Lotta Römhildt, Elke Boschke, Larysa Baraban, Jörg Opitz, Gianaurelio Cuniberti
Deutsche Biosensorsymposium 2013
For biosensor applications the choice of receptors is decisive for their sensitivity and specificity to the target. To obtain ligands for various implementations, bacteriophages have shown to possess a great potential. Within this group of bacteria infecting viruses, the filamentous phage M13 is widely studied in phage display based systems. Peptides, small proteins and antibody fragments can be presented as a fusion to phage coatproteins. These molecules and even the whole phages can be used as receptor units in biosensing. Compared to standard detecting agents like antibodies, they are more stable under varying conditions and their small size can lead to a higher sensitivity in many kinds of novel sensors.
In a selection process called biopanning, ligands for a distinct target (e.g. norovirus-like particles) can be identified using a library of phage-displayed peptides or proteins . This powerful technique, first introduced in 1985 by G. Smith , has lead to many applications in bioscience and medicine. However, there is always the risk to select target-unrelated phages . Since biopanning is a multistep procedure and dependent on a number of parameters like choice of library, immobilization and blocking strategy as well as washing and elution conditions, several options for optimizing and trouble-shooting of this enrichment process will be presented and discussed here. Subsequent to biopanning selections, different assays were established and will be compared to evaluate the binding capability and specificity of phage clones to the chosen target.
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Prof. Dr. Gianaurelio Cuniberti
Institute for Materials Science
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01062 Dresden, Germany