title: High-Performance Liquid Chromatography and Mass Spectrometry-Based Design of Proteolytically Stable Antimicrobial Peptides.
abstract: The emergence of multiresistant bacteria worldwide together with the shortage of effective antibiotics in the market emphasizes the need for the design and development of the promising agents for the treatment of superbug-associated infections. Antimicrobial peptides (AMPs) have been considered as excellent candidates to tackle this issue, and thousands of peptides of different lengths, amino acid compositions, and mode of action have been discovered and prepared to date. Nevertheless, it is of great importance to develop innovative formulation strategies for delivering these AMPs and to improve their low bioavailability and metabolic stability, particularly against proteases, if these peptides are to find applications in the clinic and administered orally or parenterally or used as dietary supplements. The purpose of this chapter is to describe basic experimental principles, based on analytical reversed-phase high-performance liquid chromatography (RP-HPLC) and mass spectrometry (MS), for the prospective design of orally bioavailable AMPs considering the structural characteristics of the peptides and the substrate specificity of proteases that abound in the body especially at sites of infection.
authors.affiliation: Peptide Chemistry Laboratory, Institute of Biochemistry and Biophysics, University of Tehran, 16 Azar Street, 1417614335, Tehran, Iran. mojtabagheri@ut.ac.ir.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Bagheri
authors.affiliation: Department of Microbiology and Immunology, Center forMicrobial Diseases and Immunity Research, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Robert E W
authors.initials: RE
authors.lastname: Hancock
mesh_terms: D000595:Amino Acid Sequence; D000890:Anti-Infective Agents / Q000737:chemistry*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase; D015195:Drug Design; D004355:Drug Stability; D006868:Hydrolysis; D013058:Mass Spectrometry / Q000379:methods; D015394:Molecular Structure; D010447:Peptide Hydrolases / Q000737:chemistry; D059748:Proteolysis; D013379:Substrate Specificity
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D010447:Peptide Hydrolases
keywords: Antimicrobial peptides; Bioavailability; Mass spectrometry; Proteases; Reversed-phase high-performance liquid chromatography