Expanding the chemical diversity of M13 bacteriophage

Abstract

Bacteriophage M13 virions are very stable nanoparticles that can be modified by chemical and genetic methods. The capsid proteins can be functionalized in a variety of chemical reactions without loss of particle integrity. In addition, Genetic Code Expansion (GCE) permits the introduction of non-canonical amino acids (ncAAs) into displayed peptides and proteins. The incorporation of ncAAs into phage libraries has led to the discovery of high-affinity binders with low nanomolar dissociation constant (KD) values that can potentially serve as inhibitors. This article reviews how bioconjugation and the incorporation of ncAAs during translation have expanded the chemistry of peptides and proteins displayed by M13 virions for a variety of purposes.

Smartphone-read phage lateral flow assay for point-of-care detection of infection

“…Thus, it is the preferred diagnostic target (over the mutation-prone surface trimer spike protein) 39 and the majority of the SARS-CoV-2 antigen tests under FDA EUA target the nucleoprotein. 40 We initially investigated different strategies for the direct conjugation of anti-nucleoprotein antibodies and HRP enzymes on the primary amines of the phage proteins, 41 including the commonly-used carbodiimide-mediated crosslinking with EDC and sulfo-NHS, Traut's reagent thiolation of primary amines for coupling to maleimide-activated antibodies, 23,24 and periodate-mediated oxidation of glycosylated antibody Fc and HRP sugars to amine-reactive aldehydes. All antibody-HRP phage conjugates tested performed well in ELISA and were captured on LFA control lines but the signal for nucleoprotein on test line was not detectable.…”

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

“…Major capsid protein pVIII consists of about 2700 copies, which accounts for 98% of the whole mass of M13 phage and contains 50 amino acids (AA). Each pVIII is composed of three domains: positively charged domain (40-50 AA) that interacts electrostatically with phage genomic DNA, intermediate hydrophobic domain (21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39), and the N-terminal domain (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20). A total of 2700 copies of pVIII protein are helically wrapped around phage DNA through electrostatic interaction between phage genomic DNA and the positively charged domain of pVIII protein, leaving the M13 surface with negative charge.…”

“…As the same kind of reactive group may exist in more than one type of amino acid residues on M13 capsid protein, cross-reactions would inevitably occur, which would greatly reduce the modification efficiency of the desired sites. To realize site-specific modification, non-canonical amino acids (ncAAs) can be integrated into the capsid protein of M13 phage [36,37]. Selenocysteine (Sec), the 21st amino acid, was displayed on the pIII protein of M13 phage using a natural selenocysteine opal suppressing tRNA.…”

Advancements in High‐Throughput Screening and Machine Learning Design for 2D Ferromagnetism: A Comprehensive Review

“…The biggest advantage of neural network-based deep learning algorithms over traditional machine learning algorithms in novel material prediction lies in their stronger learning ability and generalization capability. [123][124][125][126][127][128] The search for 2D metal-organic framework (MOF) materials with substantial perpendicular MAE is of great importance in the development of high-density magnetic storage systems. Furthermore, with the advancement of machine learning algorithms, deep learning approaches, including neural networks, have found numerous applications in the exploration of 2DFM materials.…”