A technology for developing synbodies with antibacterial activity.
The rise in antibiotic resistance has led to an increased research focus on discovery of new antibacterial candidates. While broad-spectrum antibiotics are widely pursued, there is evidence that resistance arises in part from the wide spread use of these antibiotics. Our group has developed a system to produce protein affinity agents, called synbodies, which have high affinity and specificity for their target. In this report, we describe the adaptation of this system to produce new antibacterial candidates towards a target bacterium. The system functions by screening target bacteria against an array of 10,000 random sequence peptides and, using a combination of membrane labeling and intracellular dyes, we identified peptides with target specific binding or killing functions. Binding and lytic peptides were identified in this manner and in vitro tests confirmed the activity of the lead peptides. A peptide with antibacterial activity was linked to a peptide specifically binding Staphylococcus aureus to create a synbody with increased antibacterial activity. Subsequent tests showed that this peptide could block S. aureus induced killing of HEK293 cells in a co-culture experiment. These results demonstrate the feasibility of using the synbody system to discover new antibacterial candidate agents.
The Biodesign Institute of Arizona State University, Tempe, Arizona, United States of America.
Valeriy
V
Domenyuk
Andrey
A
Loskutov
Stephen Albert
SA
Johnston
Chris W
CW
Diehnelt
D000900:Anti-Bacterial Agents / Q000138:chemical synthesis* / Q000494:pharmacology; D001412:Bacillus subtilis / Q000187:drug effects* / Q000254:growth & development; D018920:Coculture Techniques; D004926:Escherichia coli / Q000187:drug effects* / Q000254:growth & development; D005456:Fluorescent Dyes; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008826:Microbial Sensitivity Tests; D019151:Peptide Library; D040081:Protein Array Analysis; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000254:growth & development; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000254:growth & development
D000900:Anti-Bacterial Agents; D005456:Fluorescent Dyes; D019151:Peptide Library