title: Recombinant antimicrobial peptides efficiently produced using novel cloning and purification processes.
abstract: Endogenous antimicrobial peptides are ubiquitous components of animal and plant host defences. These peptides, usually cationic and amphipathic, kill target cells rapidly and are efficacious against antibiotic-resistant and clinically relevant pathogens. A practical challenge in the development of cationic peptides as therapeutics is to meet the production requirements for large quantities of highly purified drug substance at competitive costs. While chemical peptide synthesis can be used to manufacture cationic peptides, we have developed cost-effective methods for recombinant production by expressing fusion proteins comprised of multiple copies of the peptides. The fusion proteins accumulate in Escherichia coli inclusion bodies and constitute over 50% of the total cellular proteins. Active antimicrobial peptides are released by chemical reagents and purified by chromatography, combining both standard and novel approaches. Challenges of industrial-scale manufacturing of therapeutics were considered in the development of this process.
authors.affiliation: Micrologix Biotech Inc., 3650 Wesbrook Mall, Vancouver, BC, Canada.
authors.forename: Luba
authors.initials: L
authors.lastname: Metlitskaia
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Cabralda
authors.forename: Dinar
authors.initials: D
authors.lastname: Suleman
authors.forename: Cynthia
authors.initials: C
authors.lastname: Kerry
authors.forename: Jacquelyn
authors.initials: J
authors.lastname: Brinkman
authors.forename: Daniel
authors.initials: D
authors.lastname: Bartfeld
authors.forename: M Marta
authors.initials: MM
authors.lastname: Guarna
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000627:therapeutic use; D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification* / Q000627:therapeutic use; D002498:Centrifugation; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002854:Chromatography, Paper; D003001:Cloning, Molecular; D004526:Efficiency; D019075:Electrophoresis, Capillary; D004926:Escherichia coli / Q000166:cytology; D005612:Freeze Drying; D002479:Inclusion Bodies / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis; D011993:Recombinant Fusion Proteins / Q000191:economics / Q000302:isolation & purification*; D040141:Staphylococcaceae / Q000187:drug effects; D013211:Staphylococcus aureus / Q000187:drug effects
chemical_list: D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D011993:Recombinant Fusion Proteins; C088386:homoserine lactone; D015107:4-Butyrolactone