The surge in mortality and morbidity rates caused by multidrug-resistant (MDR) bacteria prompted a renewal of interest bacteriophages (phages) as clinical therapeutics natural biocontrol agents. Nevertheless, phages are continually under the pressure evolutionary phage–host arms race for survival, which is mediated co-evolving resistance mechanisms. In Anderson phage typing scheme Salmonella Ty...

Substrate identification is the key to defining molecular pathways or cellular processes regulated by proteases. Although phage display with random peptide libraries has been used to analyze substrate specificity of proteases, it is difficult to deduce endogenous substrates from mapped peptide motifs. Phage display with conventional cDNA libraries identifies high percentage of non-open reading ...

We have reported variants of the M13 bacteriophage major coat protein (P8) that enable high copy display of monomeric and oligomeric proteins, such as human growth hormone and steptavidin, on the surface of phage particles (Sidhu SS, Weiss GA, Wells JA. 2000. High copy display of large proteins on phage for functional selections. J Mol Biol 296:487-495). Here, we explore how an optimized P8 var...

An Achilles heel inherent to all molecular display formats, background binding between target and display system introduces false positives into screens and selections. For example, the negatively charged surfaces of phage, mRNA, and ribosome display systems bind with unacceptably high nonspecificity to positively charged target molecules, which represent an estimated 35% of proteins in the hum...

Despite the fact that a multitude of antibody phage display libraries has been built, systematic comparisons of critical design parameters are rare. Here we analysed the impact of various factors on the performance of the phage display system. First, we compared several vector designs for the display of Fab fragments of antibodies. Bicistronic as well as monocistronic expression of the antibody...

Phage display has been used as a high-throughput platform for identifying proteins or peptides with desired binding or catalytic activities from a complex proteome. Recently, phage display has been applied to profile the catalytic activities of posttranslational modification (PTM) enzymes. Here, we highlight recent work elucidating the downstream targets of PTM enzymes by phage display, includi...

Identification of novel vaccine candidate antigens using phage display technology J. Newcombe and J. McFadden SBLS University of Surrey, Guildford, Surrey, GU2

Peptides derived from phage display typically show significantly weaker binding than their respective high affinity phage, which can bind to protein surfaces in a multivalent fashion. Here we show that mimicking key aspects of the multivalent architecture of the phage on an AB(5) dendritic wedge can enhance the affinity of a phage-display derived collagen binding peptide 100-fold (K(d) = 550 nM...

The display of peptides and proteins on the surface of filamentous bacteriophage is a powerful methodology for selection of peptides and protein domains, including antibodies. An advantage of this methodology is the direct physical link between the phenotype and the genotype, as an analyzed polypeptide and its encoding DNA fragment exist in one phage particle. Development of phage display antib...