Affinity Selection of Peptide Binders with Magnetic Beads &lt;i&gt;via&lt;/i&gt; Organic Phase Separation (MOPS)

Murai, Ryuichi; Nogi, Taiki; Tateoka, Komei; Sato, Atsushi

doi:10.1248/bpb.b15-00455

Cited by 4 publications

(2 citation statements)

References 12 publications

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“…The generation of a phage-displayed random peptide library allows, through biopanning, the isolation of peptides that will bind to target molecules by binding affinity. The biopanning strategy consists of: (i) immobilizing the target (e.g., antibody for antigen identification) on a solid support [ 219 ], solution phase [ 220 ], or magnetic beads via organic phase separation [ 221 ]; (ii) incubating a population of phage particles with a random peptide or protein for some time, allowing the interaction between the phage-displayed peptide/protein and the target; (iii) a series of washes to remove unbound phages; (iv) elution of the phages that have maintained binding using strategies capable of breaking the bindings but maintaining phage infectivity; and (v) amplification of the eluated phages through bacterial infection. Typically, three to five rounds of affinity selection are required to reach the most strongly binding ligands, followed by isolation, enrichment, and characterization by DNA sequencing [ 215 , 222 , 223 ].…”

Section: Biotechnological Applicationsmentioning

confidence: 99%

Bacteriophages as Biotechnological Tools

Elois

Silva

Pilati

et al. 2023

Viruses

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Bacteriophages are ubiquitous organisms that can be specific to one or multiple strains of hosts, in addition to being the most abundant entities on the planet. It is estimated that they exceed ten times the total number of bacteria. They are classified as temperate, which means that phages can integrate their genome into the host genome, originating a prophage that replicates with the host cell and may confer immunity against infection by the same type of phage; and lytics, those with greater biotechnological interest and are viruses that lyse the host cell at the end of its reproductive cycle. When lysogenic, they are capable of disseminating bacterial antibiotic resistance genes through horizontal gene transfer. When professionally lytic—that is, obligately lytic and not recently descended from a temperate ancestor—they become allies in bacterial control in ecological imbalance scenarios; these viruses have a biofilm-reducing capacity. Phage therapy has also been advocated by the scientific community, given the uniqueness of issues related to the control of microorganisms and biofilm production when compared to other commonly used techniques. The advantages of using bacteriophages appear as a viable and promising alternative. This review will provide updates on the landscape of phage applications for the biocontrol of pathogens in industrial settings and healthcare.

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Section: Biotechnological Applicationsmentioning

confidence: 99%

Bacteriophages as Biotechnological Tools

Elois

Silva

Pilati

et al. 2023

Viruses

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show abstract

“…The removal of unbound phages and recovery of target-bound phages are performed through centrifugation. This methodology that applies the concept of organic phase separation to the affinity selection of phage display libraries is called magnetic beads via organic phase separation (MOPS) [ 28 ]. MOPS has been demonstrated to present high efficacy in selecting binders to Fc-fusion constructs and improve the isolation of specific binders for soluble protein targets that could not be enriched by conventional biopanning.…”

Section: Biopanning For Screening Of Combinatorial Phage Display Librmentioning

confidence: 99%

Phage display as a promising approach for vaccine development

et al. 2016

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Bacteriophages are specific antagonists to bacterial hosts. These viral entities have attracted growing interest as optimal vaccine delivery vehicles. Phages are well-matched for vaccine design due to being highly stable under harsh environmental conditions, simple and inexpensive large scale production, and potent adjuvant capacities. Phage vaccines have efficient immunostimulatory effects and present a high safety profile because these viruses have made a constant relationship with the mammalian body during a long-standing evolutionary period. The birth of phage display technology has been a turning point in the development of phage-based vaccines. Phage display vaccines are made by expressing multiple copies of an antigen on the surface of immunogenic phage particles, thereby eliciting a powerful and effective immune response. Also, the ability to produce combinatorial peptide libraries with a highly diverse pool of randomized ligands has transformed phage display into a straightforward, versatile and high throughput screening methodology for the identification of potential vaccine candidates against different diseases in particular microbial infections. These libraries can be conveniently screened through an affinity selection-based strategy called biopanning against a wide variety of targets for the selection of mimotopes with high antigenicity and immunogenicity. Also, they can be panned against the antiserum of convalescent individuals to recognize novel peptidomimetics of pathogen-related epitopes. Phage display has represented enormous promise for finding new strategies of vaccine discovery and production and current breakthroughs promise a brilliant future for the development of different phage-based vaccine platforms.

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