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WIREs Nanomed Nanobiotechnol
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Phage nanofibers in nanomedicine: Biopanning for early diagnosis, targeted therapy, and proteomics analysis

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Abstract Display of a peptide or protein of interest on the filamentous phage (also known as bacteriophage), a biological nanofiber, has opened a new route for disease diagnosis and therapy as well as proteomics. Earlier phage display was widely used in protein–protein or antigen–antibody studies. In recent years, its application in nanomedicine is becoming increasingly popular and encouraging. We aim to review the current status in this research direction. For better understanding, we start with a brief introduction of basic biology and structure of the filamentous phage. We present the principle of phage display and library construction method on the basis of the filamentous phage. We summarize the use of the phage displayed peptide library for selecting peptides with high affinity against cells or tissues. We then review the recent applications of the selected cell or tissue targeting peptides in developing new targeting probes and therapeutics to advance the early diagnosis and targeted therapy of different diseases in nanomedicine. We also discuss the integration of antibody phage display and modern proteomics in discovering new biomarkers or target proteins for disease diagnosis and therapy. Finally, we propose an outlook for further advancing the potential impact of phage display on future nanomedicine. This article is categorized under: Biology‐Inspired Nanomaterials > Protein and Virus‐Based Structures
Schematic picture of filamentous bacteriophage
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Schematic overview of the selection of antibodies (“panning“) to the targeting protein by phage display. (Reprinted with permission from Dong et al. (). Copyright 2019 Dove Press Ltd)
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In vivo detection of small pancreatic ductal adenocarcinoma (PDAC) and precursor lesions by intravital confocal microscopy and MRI using PDAC‐homing peptide modified magneto‐fluorescent nanoparticles. (a) Three slices from an ex vivo MRI of the pancreas from a 9‐week‐old Kras/p53L/+ mouse show the nanoparticle uptake (yellow arrow), which corresponds to tumor as shown in correlated HE staining sections (b), but not to regions of ductal metaplasia or normal pancreas (labeled). LN: lymph node. The image was captured at 4× objectives. (c) Fluorescence images show the uptake of Cy5.5‐labeled PTP‐NP in tumor (left) but not in normal tissue (right). (Reprinted with permission from Kelly et al. (). Copyright 2008 Kelly et al)
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Embryonic stem (ES) cell targeting by peptide modified quantum dots. (Left) Scheme of the peptide modified quantum dots. (Right) Merged optical and fluorescent images of ES cells labeled with APWHLSSQYSRT peptide modified quantum dots. Red fluorescence signal is from quantum dots. Scale bar, 100 μm. (Reprinted with permission from Lu et al. (). Copyright 2010 Lu et al)
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Photothermal breast cancer therapy using the targeting peptide modified gold nanorods. (a, b) the breast tumor image and volume changes after the therapy. A total of four groups (n = 4) of samples and controls were treated by photothermal therapy; (c) images of hematoxylin and eosin (H&E) stained sections of breast tumors obtained 3 hr after 808 nm laser treatment. Contiguous cell necrosis (dashed ovals), karyolysis, and nuclear swelling (solid ovals), cell apoptosis with the presence of nuclear pyknosis, hepereosinophilic cytoplasm (arrows), apoptotic bodies (arrow heads), and the normal tumor tissue area (angled dashed frames) are all marked. Reprinted with permission from Qu et al. (). Copyright 2017 Elsevier)
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Use of tumor‐homing gold nanorods for targeted photothermal therapy. (a) Image of the selected fluorescently labeled peptide at the MCF‐7 tumors by an in vivo fluorescence imaging system; (b) After conjugated with the best tumor‐homing peptide, the gold nanorods (AuNRs) could be enriched in the breast tumor compared to the nontargeting control groups. (Reprinted with permission from Qu et al. (). Copyright 2017 Elsevier)
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Photodynamic therapy (PDT) targeting melanoma tumor using the photosensitizer (PS)‐conjugated phage. (a) The Schematic of in vivo/in vitro biopanning was employed to select the tumor‐homing peptide. (b) The schematic of the preparation of the biomedicine based on phage and the subsequent cancer therapy. (c) The tumor volume changes after photodynamic therapy (PDT) of the biomedicine therapy. Peptide sequences: C2: SGLYKVAYDWQH, C1: GLHTSNTNLYLH, T1: AYPQKFNNNFMS, C3: SQDIRTWNGTRS (Reprinted with permission from Yang et al. (). Copyright 2018 Royal Society of Chemistry)
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Use of phage nanofibers for targeted photodynamic cancer therapy. (a) Schematic of the construction of dually functionalized phage via a chemical conjugation of pyropheophorbid‐a partially onto the genetically engineered phage that surface‐displays the peptide VSSTQDF for developing targeted photodynamic therapy. (b) in vitro treatment of MCF‐7 cells (control, B‐1, B‐2, and B‐3) and target SKBR‐3 cells (B‐4, B5, and B‐6) by targeted photodynamic therapy: (B‐1 and B‐4) optical microscopy of MCF‐7 and SKBR‐3 cells; (B‐2 and B‐5) fluorescent images of MCF‐7 and SKBR‐3 cells upon incubation with pyropheophorbid‐a and irradiation using a cell viability kit; (B‐3 and B‐6) fluorescent images of MCF‐7 and SKBR‐3 cells upon incubation with dually functionalized phage and irradiation using a cell viability kit. All images were captured at 10× objectives. (Reprinted with permission from Gandra et al. (). Copyright 2012 Wiley)
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Evaluation of the treatment of ovarian tumor by HO8910 cancer cell‐binding peptide, SWQIGGN, in nude mice. (a) Abdominal circumference measurements of three nude mice: peptide 1: SWQIGGN. (b) Tumors after the treatment of peptide 1, control peptide, and nontreatment. (Reprinted with permission from Zhou, Kang, et al. (). Copyright 2015 Zhou et al.)
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Cytotoxicity of MCF‐7‐targeted phage‐micelles against MCF‐7 cells 72 hr after treatment. Controls include paclitaxel (PCT)‐loaded plain micelles, irrelevant SA‐phage‐micelles, drug‐free MCF‐7‐targeted phage‐micelles, free PCT in dimethylsulfoxide (DMSO), and drug‐free SA‐phage‐micelles (*p < .05, mean ± SD, n = 6). (Reprinted with permission from Wang, Petrenko, and Torchilin (). Copyright 2010 American Chemical Society)
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In vivo localization of fluorescent (Alexa 488) CKS9 peptide modified chitosan nanoparticles (CKS9‐CNs) on the rat Peyer's patch region 1 hr after injection. (a) CNs‐Alexa 488, (b) CKS9‐CNs‐Alexa 488. Green fluorescent signal indicates the presence of chitosan nanoparticles. Red fluorescent signal indicates the presence of follicle‐associated epithelium (FAE). (Reprinted with permission from Yoo et al. (). Copyright 2010 Elsevier)
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Fas gene silencing by cardiomyocyte‐targeted primary cardiomyocyte‐modified bioreducible polymer. (a) Fas expression evaluation in H9C2 cells transfected by Fas siRNA under normoxic or hypoxic conditions measured by flow cytometry. (b) The percentage of Fas expressing H9C2 cells. (c) The Fas expression level under hypoxia (normalized to control non‐treated H9C2 cells, 100%). * indicates p < .05. (Reprinted with permission from Nam, McGinn, Kim, Kim, and Bull (). Copyright 2010 Elsevier)
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Use of phage‐derived rat mesenchymal stem cells (rMSC)‐targeting major coat protein (pVIII) for enhanced gene delivery to rMSCs. (a) Scheme for functionalizing magnetic silica nanoclusters with rMSC‐binding pVIII for gene delivery to rMSCs; (b) enhanced green fluorescent protein–vascular endothelial growth factor (EGFP–VEGF) (EGFP tagged vascular endothelial growth factor) gene transfection efficiency by magnetic silica nanoclusters and controls. (c) Transfection expression of EGFP mediated by Lipofectamine2000 (a), LPD (b), and LPD with peptide 1 (c), peptide 2 (d), peptide 3 (e), peptide 1 + 2 (f) in rMSCs using the EGFP reporter. Image 1 shows EGFP expression and image 2 shows all the cells in the same area. Scale bar: 100 μm. CTAB, cetyltrimethylammonium bromide; MPTS, (3‐mercaptopropyl) trimethoxysilane; PEI, polyethylenimine; WT, wild type. (Reprinted with permission from Ma, Wang, et al. (). Copyright 2013 Wiley)
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The procedure of phage biopanning against cells in vitro or tissues in vivo
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