Construction of helper plasmid-mediated dual-display phage for autoantibody screening in serum

Rajaram, Kaushik; Vermeeren, Veronique; Somers, Klaartje; Somers, Veerle; Michiels, Luc

doi:10.1007/s00253-014-5713-8

Cited by 4 publications

(3 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The first one, alkaline phosphatase, or green fluorescent protein (GFP), was located on the head, and the other, anti-CEA scFv, on the tail of the lambda phage [ 237 ]. A similar approach was used by Rajaram et al, who also created a dual-display system [ 238 ] in which a biomarker and a short peptide containing the streptavidin-binding motif Histidine-Proline-Glutamine (HPQ) were displayed on the opposite ends of the M13 bacteriophage capsid. This additional second peptide with a high affinity to streptavidin was introduced to allow for sample (phage) purification on streptavidin-conjugated magnetic beads to increase assay sensitivity and decrease background signal.…”

Section: Intranasal Administrationmentioning

confidence: 99%

Phage Interactions with the Nervous System in Health and Disease

Jędrusiak,

Fortuna,

Majewska

et al. 2023

Cells

View full text Add to dashboard Cite

The central nervous system manages all of our activities (e.g., direct thinking and decision-making processes). It receives information from the environment and responds to environmental stimuli. Bacterial viruses (bacteriophages, phages) are the most numerous structures occurring in the biosphere and are also found in the human organism. Therefore, understanding how phages may influence this system is of great importance and is the purpose of this review. We have focused on the effect of natural bacteriophages in the central nervous system, linking them to those present in the gut microbiota, creating the gut-brain axis network, as well as their interdependence. Importantly, based on the current knowledge in the field of phage application (e.g., intranasal) in the treatment of bacterial diseases associated with the brain and nervous system, bacteriophages may have significant therapeutic potential. Moreover, it was indicated that bacteriophages may influence cognitive processing. In addition, phages (via phage display technology) appear promising as a targeted therapeutic tool in the treatment of, among other things, brain cancers. The information collected and reviewed in this work indicates that phages and their impact on the nervous system is a fascinating and, so far, underexplored field. Therefore, the aim of this review is not only to summarize currently available information on the association of phages with the nervous system, but also to stimulate future studies that could pave the way for novel therapeutic approaches potentially useful in treating bacterial and non-bacterial neural diseases.

show abstract

Section: Intranasal Administrationmentioning

confidence: 99%

Phage Interactions with the Nervous System in Health and Disease

Jędrusiak,

Fortuna,

Majewska

et al. 2023

Cells

View full text Add to dashboard Cite

show abstract

“…As with PhoPhabs, 44 the physical separation of genes encoding fusion proteins on two separate vectors (i.e., phagemid plus plasmid/designed helper phage) allows for simple combination of various binding and detection components on phage scaffold without the need for laborious genetic engineering of each individual probe. Another example of trans modular phage biosensor design was reported by Rajaram et al 46 They prepared filamentous virions concomitantly displaying a short streptavidin-binding peptide on p7 expressed from a helper plasmid, and phagemid-encoded UH-RA.21, a 28-amino-acid rheumatoid arthritis autoantigen, fused to p6. The helper plasmid lacked a packaging signal; therefore, recombinant phages were produced only upon transduction of host cells with phagemid virions.…”

Section: Bipartite Phage Display: Combining Twomentioning

confidence: 99%

Bacteriophages as Scaffolds for Bipartite Display: Designing Swiss Army Knives on a Nanoscale

Molek

Bratkovič

2015

Bioconjugate Chem.

View full text Add to dashboard Cite

Bacteriophages have been exploited as cloning vectors and display vehicles for decades owing to their genetic and structural simplicity. In bipartite display setting, phage takes on the role of a handle to which two modules are attached, each endowing it with specific functionality, much like the Swiss army knife. This concept offers unprecedented potential for phage applications in nanobiotechnology. Here, we compare common phage display platforms and discuss approaches to simultaneously append two or more different (poly)peptides or synthetic compounds to phage coat using genetic fusions, chemical or enzymatic conjugations, and in vitro noncovalent decoration techniques. We also review current reports on design of phage frameworks to link multiple effectors, and their use in diverse scientific disciplines. Bipartite phage display had left its mark in development of biosensors, vaccines, and targeted delivery vehicles. Furthermore, multifunctionalized phages have been utilized to template assembly of inorganic materials and protein complexes, showing promise as scaffolds in material sciences and structural biology, respectively.

show abstract

“…21 as a biomarker and streptavidin (STV)-binding protein (SBP) as capturing peptide, both being displayed at opposite ends of a filamentous phage. 12 This approach resulted in a higher sensitivity of signal detection assays using STV-coated magnetic beads. Specifically, the dual phage displayed a biomarker (autoantigenic target UH-RA.21 specific for rheumatoid arthritis) fused to a coat protein pVI at one end and an anchoring SBP fused to a coat protein pVII at the other end.…”

mentioning

confidence: 99%

Real-time analysis of dual-display phage immobilization and autoantibody screening using quartz crystal microbalance with dissipation monitoring

Rajaram¹,

Losada-Pérez²,

Vermeeren³

et al. 2015

IJN

Self Cite

View full text Add to dashboard Cite

Over the last three decades, phage display technology has been used for the display of target-specific biomarkers, peptides, antibodies, etc. Phage display-based assays are mostly limited to the phage ELISA, which is notorious for its high background signal and laborious methodology. These problems have been recently overcome by designing a dual-display phage with two different end functionalities, namely, streptavidin (STV)-binding protein at one end and a rheumatoid arthritis-specific autoantigenic target at the other end. Using this dual-display phage, a much higher sensitivity in screening specificities of autoantibodies in complex serum sample has been detected compared to single-display phage system on phage ELISA. Herein, we aimed to develop a novel, rapid, and sensitive dual-display phage to detect autoantibodies presence in serum samples using quartz crystal microbalance with dissipation monitoring as a sensing platform. The vertical functionalization of the phage over the STV-modified surfaces resulted in clear frequency and dissipation shifts revealing a well-defined viscoelastic signature. Screening for autoantibodies using antihuman IgG-modified surfaces and the dual-display phage with STV magnetic bead complexes allowed to isolate the target entities from complex mixtures and to achieve a large response as compared to negative control samples. This novel dual-display strategy can be a potential alternative to the time consuming phage ELISA protocols for the qualitative analysis of serum autoantibodies and can be taken as a departure point to ultimately achieve a point of care diagnostic system.

show abstract

Construction of helper plasmid-mediated dual-display phage for autoantibody screening in serum

Cited by 4 publications

References 24 publications

Phage Interactions with the Nervous System in Health and Disease

Phage Interactions with the Nervous System in Health and Disease

Bacteriophages as Scaffolds for Bipartite Display: Designing Swiss Army Knives on a Nanoscale

Real-time analysis of dual-display phage immobilization and autoantibody screening using quartz crystal microbalance with dissipation monitoring

Contact Info

Product

Resources

About