M13 phage: a versatile building block for a highly specific analysis platform

Abstract

Viruses are changing the biosensing and biomedicine landscape due to their multivalency, orthogonal reactivities, and responsiveness to genetic modifications. As the most extensively studied phage model for constructing a phage display library, M13 phage has received much research attention as building blocks or viral scaffolds for various applications including isolation/separation, sensing/probing, and in vivo imaging. Through genetic engineering and chemical modification, M13 phages can be functionalized into a multifunctional analysis platform with various functional regions conducting their functionality without mutual disturbance. Its unique filamentous morphology and flexibility also promoted the analytical performance in terms of target affinity and signal amplification. In this review, we mainly focused on the application of M13 phage in the analytical field and the benefit it brings. We also introduced several genetic engineering and chemical modification approaches for endowing M13 with various functionalities, and summarized some representative applications using M13 phages to construct isolation sorbents, biosensors, cell imaging probes, and immunoassays. Finally, current issues and challenges remaining in this field were discussed and future perspectives were also proposed.

Keywords: Bioimaging; Biosensors; Immunoassays/ELISA; M13 phage; Phage display.

Fig. 1

A Illustration of the structure of M13 phage and the amino acid sequence of pVIII protein. B Schematic showing the infection and production process of natural M13 phage. C Illustration of the mechanism of phage vector-based display system. D Illustration of the mechanism of phagemid-based display system

Fig. 2

Common chemical modifications of the inherent functional groups presented on the major coat protein of M13 phage

Fig. 3

Microfluidic-based biopanning strategies for the screening of target M13 bacteriophages. A Mechanism of droplet microfluidic-based double monoclonal display (dm-display) system. Copyright 2021 Royal Society of Chemistry [52]. B Scheme of the SCOPE system-based phage screening for micropillar array and particle tracing. Copyright 2022 National Academy of Science [53]. C Schematic illustration of the μCellect methodology. Copyright 2022 American Chemical Society [54]

Fig. 4

Representative examples of M13-based recognition strategies. A Graphic of the biopanning procedure to obtain the Cr(III) binding phages. Copyright 2015 American Chemical Society [58]. B Chimeric M13 phage decorated with AuNPs for detection of bacteria. Copyright 2019 American Chemical Society [71]. C Construction of TC-phage-FlAsH structure for bacterial detection. TC-tag displaying M13 phage recognized and infected the host bacteria and then labeled with the fluorescent dye FlAsH-EDT₂. Copyright 2014 Wiley–VCH GmbH [72]. D Schematic illustration of PhaNGS approach. The phage particles were designed to specifically binding to target proteins and identified by unique sequence through NGS. Copyright 2018 National Academy of Science [78]

Fig. 5

Optical sensors for specifical determination of biomarkers based on M13 phage particles. A Construction of volatile chemical biosensor inspired by the color-shift characteristic of turkey skin. TNT specific engineered M13 phage could self-assemble into the structure of mimic collagen fibers, which produces different colors through swelling or deswelling to indicate the concentration of TNT in the air. Copyright 2014 Springer Nature [80]. B Construction of M-phage-DNAzymes for miRNA quantification. The HRP-mimic hemin/G-quadruplex DNAzyme could catalyze the oxidation of ABTS²⁻ under the C HCR reaction triggered by target miRNA. Copyright 2022 Wiley–VCH GmbH [86]

Fig. 6

Dual functional M13 phage for pathogen capturing and sensing. A Fabrication of dynamic deformable nanointerface using flexible M13 phage and biotinylated-aptamer. B Procedure of bacterial capturing and sensing. C Schematic illustration of the significantly enhanced binding affinity and accelerated mass transport through sway motion of flexible M13 phage. Copyright 2022 Wiley-VCH GmbH [94]

Fig. 7

A Illustration of the synthesis of E3-Cy3-AgNPs complex (upper) and the super resolution fluorescence images of E. coli stained with the E3-CTPEG2K-Cy3-AgNPs probe (below). Scale bar is 1 μm. Copyright 2019 Wiley–VCH GmbH [99]. B Structure of SWNT-based M13 phage (upper) and the imaging examples of E. coli-infected mice with SWNT probe (below). Copyright 2014 Springer Nature [104]