title: Panning of a phage display library against a synthetic capsule for peptide ligands that bind to the native capsule of Bacillus anthracis.
abstract: Bacillus anthracis is the causative agent of anthrax with the ability to not only produce a tripartite toxin, but also an enveloping capsule comprised primarily of γ-D-glutamic acid residues. The purpose of this study was to isolate peptide ligands capable of binding to the native capsule of B. anthracis from a commercial phage display peptide library using a synthetic form of the capsule consisting of 12 γ-D-glutamic acid residues. Following four rounds of selection, 80 clones were selected randomly and analysed by DNA sequencing. Four clones, each containing a unique consensus sequence, were identified by sequence alignment analysis. Phage particles were prepared and their derived 12-mer peptides were also chemically synthesized and conjugated to BSA. Both the phage particles and free peptide-BSA conjugates were evaluated by ELISA for binding to encapsulated cells of B. anthracis as well as a B. anthracis capsule extract. All the phage particles tested except one were able to bind to both the encapsulated cells and the capsule extract. However, the peptide-BSA conjugates could only bind to the encapsulated cells. One of the peptide-BSA conjugates, with the sequence DSSRIPMQWHPQ (termed G1), was fluorescently labelled and its binding to the encapsulated cells was further confirmed by confocal microscopy. The results demonstrated that the synthetic capsule was effective in isolating phage-displayed peptides with binding affinity for the native capsule of B. anthracis.
authors.affiliation: Human Protection and Performance Division, Defence Science and Technology Organisation, Fishermans Bend, Victoria, Australia.
authors.forename: Michael
authors.initials: M
authors.lastname: Beer
authors.forename: Chun-Qiang
authors.initials: CQ
authors.lastname: Liu
mesh_terms: D001408:Bacillus anthracis / Q000378:metabolism*; D016667:Bacterial Capsules / Q000737:chemistry / Q000378:metabolism*; D001435:Bacteriophages / Q000378:metabolism; D008024:Ligands; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D008024:Ligands; D019151:Peptide Library; D010455:Peptides