title: Combinatorial Library Screening with Liposomes for Discovery of Membrane Active Peptides.
abstract: Membrane active peptides (MAPs) represent a class of short biomolecules that have shown great promise in facilitating intracellular delivery without disrupting cellular plasma membranes. Yet their clinical application has been stalled by numerous factors: off-target delivery, a requirement for high local concentration near cells of interest, degradation en route to the target site, and in the case of cell-penetrating peptides, eventual entrapment in endolysosomal compartments. The current method of deriving MAPs from naturally occurring proteins has restricted the discovery of new peptides that may overcome these limitations. Here, we describe a new branch of assays featuring high-throughput functional screening capable of discovering new peptides with tailored cell uptake and endosomal escape capabilities. The one-bead-one-compound (OBOC) combinatorial method is used to screen libraries containing millions of potential MAPs for binding to synthetic liposomes, which can be adapted to mimic various aspects of limiting membranes. By incorporating unnatural and d-amino acids in the library, in addition to varying buffer conditions and liposome compositions, we have identified several new highly potent MAPs that improve on current standards and introduce motifs that were previously unknown or considered unsuitable. Since small variations in pH and lipid composition can be controlled during screening, peptides discovered using this methodology could aid researchers building drug delivery platforms with unique requirements, such as targeted intracellular localization.
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Randy P
authors.identifier: 0000-0001-8193-1664
authors.initials: RP
authors.lastname: Carney
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Yann
authors.initials: Y
authors.lastname: Thillier
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Zsofia
authors.initials: Z
authors.lastname: Kiss
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Amir
authors.initials: A
authors.lastname: Sahabi
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Jean Carlos
authors.initials: JC
authors.lastname: Heleno Campos
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Alisha
authors.initials: A
authors.lastname: Knudson
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Ruiwu
authors.initials: R
authors.lastname: Liu
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: David
authors.initials: D
authors.lastname: Olivos
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Mary
authors.initials: M
authors.lastname: Saunders
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.
authors.forename: Lin
authors.initials: L
authors.lastname: Tian
authors.affiliation: Department of Biochemistry and Molecular Medicine, University of California Davis , 2700 Stockton Boulevard, Sacramento, California 95817, United States.|Division of Hematology/Oncology, University of California Davis Cancer Center , Sacramento, California United States.
authors.forename: Kit S
authors.initials: KS
authors.lastname: Lam
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D002460:Cell Line; D057846:Cell-Penetrating Peptides / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D020650:Combinatorial Chemistry Techniques; D005456:Fluorescent Dyes / Q000737:chemistry; D057166:High-Throughput Screening Assays; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008081:Liposomes / Q000737:chemistry*; D008863:Microspheres; D019151:Peptide Library; D012235:Rhodamines / Q000737:chemistry; D013499:Surface Properties
chemical_list: C114557:5-carboxytetramethylrhodamine succinimidyl ester; D000596:Amino Acids; D057846:Cell-Penetrating Peptides; D005456:Fluorescent Dyes; D008081:Liposomes; D019151:Peptide Library; D012235:Rhodamines
keywords: drug delivery platforms; endosomal escape capabilities; high-throughput; liposomes; membrane-active proteins; one-bead-one-compound