Phage display in the study of infectious diseases

Abstract

Microbial infections are dependent on the panoply of interactions between pathogen and host and identifying the molecular basis of such interactions is necessary to understand and control infection. Phage display is a simple functional genomic methodology for screening and identifying protein-ligand interactions and is widely used in epitope mapping, antibody engineering and screening for receptor agonists or antagonists. Phage display is also used widely in various forms, including the use of fragment libraries of whole microbial genomes, to identify peptide-ligand and protein-ligand interactions that are of importance in infection. In particular, this technique has proved successful in identifying microbial adhesins that are vital for colonization.

Comparative characteristic of the methods of protein antigens epitope mapping

“…Існують також й бібліотеки, що відображають всю мо-лекулу, частину протеїну чи його домен [30]. Також чимало прикладів використання фаго-вого дисплею для встановлення антигенної будови протеїнів різного походження, напри-клад: α 2 -макроглобулін [39], PDZ-домени різних адаптерних протеїнів [40], вірусні протеїни (нуклеокапсидний протеїн вірусу свинячого репродуктивного та респіраторного синдрому [41], капсидний протеїн вірусу катаральної лихо-манки овець [42], глікопротеїн вірусу класичної свинячої лихоманки [43]), протеїни-антигени патогенних мікроорганізмів (гемофільної па-лички Пфейффера Haemophilus influenzae, збудника м'якого шанкру Haemophilus ducreyi, одного з етіологічних агентів парадонтиту Aggregatibacter actinomycetemcomitans, збудник пастерельозу Pasteurella multocida [44], енто-мопатогенний мікроскопічний гриб Beauveria bassiana [45]). …”

Employing phage display to study the mode of action of Bacillus thuringiensis Cry toxins

“…Most commonly, phage display libraries are constructed using vectors called phagemids, which are hybrids of phage and plasmid vectors. These phagemids contain the origins of replication from the M13 phage and E. coli; the gene coat protein III or gene coat IV for protein fusion, and an antibiotic resistance gene [1,25]. To display the fusion protein, E. coli cells harboring the phagemids are infected with a helper phage that provides all the necessary components for phage assembly.…”

“…One of the most important features of phage display is that it allows a rapid selection of variants with improved binding characteristics [1,25]. Even more, further mutagenesis and selection by panning allows in vitro molecular evolution of proteins [5].…”

Development of a cell-based fluorescence resonance energy transfer reporter for Bacillus anthracis lethal factor protease

“…The combination of FACS with FRET-based genetically encoded proteolytic reporters allows the possibility for in vivo screening of protease inhibitors using chemically generated [12; 13] or biologically encoded libraries [14]. The later possibility suggests an even more intriguing scenario in which single cells can be used as individual microfactories for the biosynthesis and screening of particular inhibitors in a single process within the same cellular cytoplasm [26].…”