Methods to screen peptide libraries using minicell display
AssigneeChildren's Medical Center Corporation
InventorSamy Ashkar
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Abstract
A minicell display method has been developed which has significant advantages for screening peptide libraries for candidates that can bind and effectively modulate a particular biological process. The method, based on the small, anucleate minicell, has increased versatility in generating unique sequences to screen as well as increasing the size of the peptides to be screened. In vivo mutagenesis, at the level of protein synthesis, as well as DNA replication, increases diversification of the library to be screened and therefore substantially increases the number of potential peptides that can modulate a particular biological response or mechanism.
Claims20
A method for identifying minicell hosts bound to a binding partner comprising:
- (a) expressing a fusion protein in a minicell host comprising an outer membrane, wherein the fusion protein is encoded by a chimeric gene comprising:
- a DNA fragment encoding an N-terminal fragment of a 17K antigen of Rickettsia rickettsii consisting essentially of a signal sequence and lipid modification site which mediates localization of the fusion protein to the outer membrane, and a DNA fragment encoding a second peptide;
- (b) contacting the minicell host of step (a) with a binding partner;
- and (c) identifying the minicell hosts bound to the binding partner.
Dependencies are inferred from the claim text.