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Review
. 2014 Jan 22;114(2):1020-81.
doi: 10.1021/cr400166n. Epub 2013 Dec 3.

Combinatorial peptide libraries: mining for cell-binding peptides

Bethany Powell Gray  1 , Kathlynn C Brown
Affiliations
Free PMC article
Review

Combinatorial peptide libraries: mining for cell-binding peptides

Bethany Powell Gray et al. Chem Rev. .
Free PMC article
No abstract available

PubMed Disclaimer

Figures

Figure 1
Figure 1
Peptide libraries used for the selection of cell-binding peptides. Biological and chemical peptide libraries have been used to isolate cell-specific peptides. For phage and bacterial display, the diversity is generated at the DNA level and there is an inherent genotype–phenotype connection. For one-bead one-compound and positional scanning synthetic peptide libraries, the diversity is generated chemically and is based on the use of a collection of monomers. The resultant peptides are displayed in red for clarity. The PS-SPCL schematic illustrates the pools of peptide libraries generated for a tetrameric peptide where each of the 20 amino acids is a unique colored circle and the mixture of 20 amino acids is shown in blue.
Figure 2
Figure 2
Filamentous and lytic phage structures. (A) Schematic representation of fd filamentous phage. The random peptide is shown fused to the amino terminus of the pIII coat protein. (B) Representative T7 lytic phage structure. The T7 phage head is comprised of the 10A and 10B capsid proteins arranged as hexamer or pentamer units at a total of 415 proteins per head. A graphical representation of the hexamer capsid unit is shown with a random peptide (red) fused to the 10B protein (blue triangle). T7 phage can be modified to express varying ratios of 10B to 10A protein, displaying peptide sequences in 1–415 copies.
Figure 3
Figure 3
Different types of bacterial display peptide libraries. Peptide libraries, including FliTrx, OmpA, CPX, and invasin libraries, have been incorporated into bacterial membrane proteins at different locations as shown in the four panels. (A) Peptide library insertion into the middle of the membrane protein. (B) Display of the peptide library at the N-terminus of the membrane protein. (C) Display of the peptide library at the C-terminus of the membrane protein. (D) Display of the peptide library through a combination of N- and C-terminal display.
Figure 4
Figure 4
OBOC peptide library generation using “split-mix” synthesis. An example of split-mix synthesis for tripeptides composed of leucine (L), alanine (A), and threonine (T) is shown. The beads are divided into three different pools, one pool for conjugation to each of the amino acids using standard solid-phase synthesis. Pool 1 is coupled to L, pool 2 to A, and pool 3 to T. The beads from all pools are combined and randomly split into three new pools before a second round of amino acid conjugation. As before, pool 1 beads are coupled to L, pool 2 beads to A, and pool 3 beads to T. Finally, the pools are mixed and randomly sorted again for another round of amino acid conjugation. This results in a library of bead-bound peptides composed of every combination of the 3 amino acids, totaling 27 different peptide sequences (33).
Figure 5
Figure 5
Design of a PS-SPCL library. Mixture 1 consists of all peptides with a first amino acid of “A”, while mixture 2 is all peptides with a first amino acid of “C”. Each of mixtures 3–20 displays 1 of the remaining 18 amino acids in the first amino acid position. The next library subset, mixtures 21–40, contains 1 of the 20 amino acids held constant in the second library position. This scanning is continued until each of the tetrapeptide positions has its own pool of libraries. This is represented graphically with each amino acid being represented by a unique colored circle and a mixture of the 20 amino acids being represented as a blue circle.
Figure 6
Figure 6
Panning of phage-displayed peptide libraries. In each case, the phage library is bound to the target, which can be a purified protein, viable cells, or an animal. Nonbinding clones are removed by stringent washes, and phage associated with the target are amplified in E. coli. The process is repeated, enriching for binding peptides at each round.
Figure 7
Figure 7
A phage-display-selected peptide inhibits tumor metastasis. B16F10-Nex 2 cells were injected intravenously into mice, and the mice were treated via an intraperitoneal injection with peptide 20 (CSSRTMHHC), a scrambled control peptide, or buffer. Images of the lungs show a reduction in metastatic nodules (brown spots) in the animals treated with peptide 20. Reprinted with permission from ref 160. Copyright 2010 Springer-Verlag.
Figure 8
Figure 8
Structures of common cleavable linkers used to attach drugs to targeting peptides. (A) Cleavable linkers are shown along with the conditions which release the drug from the peptide carrier. The acetal, ketal, and hydrazone linkers require incorporation of reactive moieties not found within the 20 naturally occurring amino acids. (B) Two types of commonly used self-immolative linkers are shown along with the mechanism in which drug is released from the carrier. The release is initiated by a trigger, such as a cleavage reaction shown in panel A, where X = O, S, or NH and Y = CH2, NR, or O. The drug serves as a leaving group and is typically attached as an ester or amide. In both cases, the peptide is represented as a squiggly line.
Figure 9
Figure 9
Vaccination with plasma membrane vehicles modified with the dendritic-cell-specific peptide CGRWSGWPADLC (p30) leads to antitumor activity. Naïve mice were injected iv with B16-OVA cells on day 0. At days 2, 8, and 14 different groups of mice (five mice per group) were vaccinated with PBS or B16-OVA-derived plasma membrane vehicles modified with the control peptide 12His or the dendritic-cell-specific peptide p30. At day 21, the lungs were removed from the mice, and tumor foci were counted via microscopy. (A) Bars indicate the mean number of tumor foci for each vaccination group, and this number is indicated above each bar. (B) Representative lung images from each vaccination group. Reprinted with permission from ref 391. Copyright 2010 Union for International Cancer Control (UICC).
Figure 10
Figure 10
The CTP peptide allows for specific cardiac imaging. Mice were imaged by in vivo fluorescent imaging following intracardiac injection of fluorospheres alone, CTP peptide–fluorospheres, or control peptide–fluorospheres. Shown are representative images from the three mice per group at different time points postinjection. Heart accumulation, indicated by the arrow, is only observed for the CTP peptide–fluorospheres. Reprinted with permission from ref 358. Copyright 2010 Public Library of Science (PLoS).
Figure 11
Figure 11
Small-animal PET/CT imaging of EphB4-positive tumors. The EphB4-binding peptide TNYLFSPNGPIARAW was conjugated to DOTA and loaded with 64Cu. Signal is observed at 4 h in CT-26 and PC-3M but not in the EphB4-negative A549 tumors. Reprinted with permission from ref 125. Copyright 2011 Society of Nuclear Medicine, Inc.
Figure 12
Figure 12
VCAM-1-specific peptide homes to areas of inflammation and atherosclerotic deposits. (A) The VCAM-1-binding peptide CVHSPNKKCGGSKGK was coupled to a magnetofluorescent nanoparticle (VPN). TNF-α-induced inflammation was induced in one ear of a mouse, while the other was left untreated. Intravital microscopy shows clear accumulation of VPN (red) in the inflamed ear at 4 h, while no binding is observed in the normal control ear. (B) Ex vivo imaging by MR and macroscopic fluorescence at 24 h post intravenous injection detects binding of VPN in animals with atherosclerotic plaques (Apo E−/− mice). No signal is detected in wild-type animals with no plaque development or when a nontargeted nanoparticle is used. The high accumulation of VPN in the aortic arch is indicated by the arrows. Adapted with permission from ref 365. Copyright 2005 American Heart Association Inc.
Figure 13
Figure 13
Structures of two multimeric peptide scaffolds. Multimeric presentation of the targeting peptides mimics the valency and orientation of the phage particle. The tetrameric peptide based on a trilysine core is shown to the left and the pentavalent dendritic wedge on the right. The targeting peptide is shown as a gray oval. A variety of chemical moieties have been attached to the trilysine core structure, indicated by “R” in the figure.
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References

    1. Strebhardt K, Ullrich A. Nat. Rev. Cancer. 2008;8:473. - PubMed
    1. Reichert JM. mAbs. 2013;5:1. - PMC - PubMed
    1. Reichert JM, Valge-Archer VE. Nat. Rev. Drug Discovery. 2007;6:349. - PubMed
    1. Tabrizi MA, Tseng C-ML, Roskos LK. Drug DiscoveryToday. 2006;11:81. - PubMed
    1. Neumeister P, Eibl M, Zinke-Cerwenka W, Scarpatetti M, Sill H, Linkesch W. Ann. Hematol. 2001;80:119. - PubMed

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