Phage Display on the Base of Filamentous Bacteriophages: Application for Recombinant Antibodies Selection

Tikunova, Nina; Morozova, Vera

doi:10.32607/20758251-2009-1-3-20-28

Cited by 36 publications

(15 citation statements)

References 85 publications

(97 reference statements)

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“…The sequences of variable genes of the TCR of the human T-cell clone T Z4B27, specific for M. tuberculosis sulfoglycolipid Ac2SGL, displayed by the CD1b molecule were obtained. To design a αβ scTCR the variable genes were linked by a flexible hydrophilic peptide consisting in glycine and serine (Gly 4 Ser) 3 , which make it flexible and resistant to proteases [6]. The phagemid pHEN1 was selected to display the scTCR linked to PIII protein of m13 phage.…”

Section: Resultsmentioning

confidence: 99%

“…Taking into account that CD1b molecules are not polymorphic, Ac 2 SGL is expressed only by virulent mycobacteria, and that lipids are not subject to mutations induced by selective pressure of the host immune system, the use of CD1b:Ac 2 SGL complex as universal marker of tuberculosis infection should be evaluated. Phage display has been instrumental for the success of antibody (Ab) technology [6], but not fully expanded to TCRs [7]. The aim of the present study was, to display a functional αβ scTCR recognizing the CD1b:Ac2SGL complex from M. tuberculosis , on the PIII protein of m13 phage.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Phage display of functional αβ single-chain T-cell receptor molecules specific for CD1b:Ac2SGL complexes from Mycobacterium tuberculosis-infected cells

Camacho

Huggett²,

Kim³

et al. 2013

BMC Immunol

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The development of molecules specific for M. tuberculosis-infected cells has important implications, as these tools may facilitate understanding of the mechanisms regulating host pathogen interactions in vivo. In addition, development of new tools capable to targeting M. tuberculosis-infected cells may have potential applications to diagnosis, treatment, and prevention of tuberculosis (TB). Due to the lack of CD1b polymorphism, M. tuberculosis lipid-CD1b complexes could be considered as universal tuberculosis infection markers. The aim of the present study was to display on the PIII surface protein of m13 phage, a human αβ single-chain T-cell receptor molecule specific for CD1b:2-stearoyl-3-hydroxyphthioceranoyl-2´-sulfate-α-α´-D-trehalose (Ac2SGL) which is a complex presented by human cells infected with M. tuberculosis. The results showed the pIII fusion particle was successfully displayed on the phage surface. The study of the recognition of the recombinant phage in ELISA and immunohistochemistry showed the recognition of CD1b:Ac2SGL complexes and cells in human lung tissue from a tuberculosis patient respectively, suggesting the specific recognition of the lipid-CD1b complex.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Phage display of functional αβ single-chain T-cell receptor molecules specific for CD1b:Ac2SGL complexes from Mycobacterium tuberculosis-infected cells

Camacho

Huggett²,

Kim³

et al. 2013

BMC Immunol

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show abstract

“…RPLs can subsequently be used in many applications including the identification of peptide ligands by receptors, the mapping of substrate sites for enzymes, and the creation of antibody peptide libraries. These applications are reviewed elsewhere [ 38 , 46 , 47 ]. Inovirus display technology has also been used for epitope mapping and vaccine design purposes.…”

Section: Inovirus Associated Vectorsmentioning

confidence: 99%

Architectural Insight into Inovirus-Associated Vectors (IAVs) and Development of IAV-Based Vaccines Inducing Humoral and Cellular Responses: Implications in HIV-1 Vaccines

Hassapis

Stylianou

Kostrikis

2014

Viruses

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Inovirus-associated vectors (IAVs) are engineered, non-lytic, filamentous bacteriophages that are assembled primarily from thousands of copies of the major coat protein gp8 and just five copies of each of the four minor coat proteins gp3, gp6, gp7 and gp9. Inovirus display studies have shown that the architecture of inoviruses makes all coat proteins of the inoviral particle accessible to the outside. This particular feature of IAVs allows foreign antigenic peptides to be displayed on the outer surface of the virion fused to its coat proteins and for more than two decades has been exploited in many applications including antibody or peptide display libraries, drug design, and vaccine development against infectious and non-infectious diseases. As vaccine carriers, IAVs have been shown to elicit both a cellular and humoral response against various pathogens through the display of antibody epitopes on their coat proteins. Despite their high immunogenicity, the goal of developing an effective vaccine against HIV-1 has not yet materialized. One possible limitation of previous efforts was the use of broadly neutralizing antibodies, which exhibited autoreactivity properties. In the past five years, however, new, more potent broadly neutralizing antibodies that do not exhibit autoreactivity properties have been isolated from HIV-1 infected individuals, suggesting that vaccination strategies aimed at producing such broadly neutralizing antibodies may confer protection against infection. The utilization of these new, broadly neutralizing antibodies in combination with the architectural traits of IAVs have driven the current developments in the design of an inovirus-based vaccine against HIV-1. This article reviews the applications of IAVs in vaccine development, with particular emphasis on the design of inoviral-based vaccines against HIV-1.

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“…This unique features enable PDT as a high-throughput Ab screening technology to be well suited for proteomic analysis in the postgenomic research era. 18 In general, such endeavors require high-throughput screening procedures via phage ELISA to identify individual phage binders. Phage ELISA is a fast and robust screening assay because it allows many clones to be screened in parallel.…”

Section: Discussionmentioning

confidence: 99%

Improved Soluble ScFv ELISA Screening Approach for Antibody Discovery Using Phage Display Technology

et al. 2017

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Phage display technology (PDT) is a powerful tool for the isolation of recombinant antibody (Ab) fragments. Using PDT, target molecule-specific phage-Ab clones are enriched through the "biopanning" process. The individual specific binders are screened by the monoclonal scFv enzyme-linked immunosorbent assay (ELISA) that may associate with inevitable false-negative results. Thus, in this study, three strategies were investigated for optimization of the scFvs screening using Tomlinson I and J libraries, including (1) optimizing the expression of functional scFvs, (2) improving the sensitivity of ELISA, and (3) preparing different samples containing scFvs. The expression of all scFv Abs was significantly enhanced when scFv clones were cultivated in the terrific broth (TB) medium at the optimum temperature of 30 °C. The protein A-conjugated with horseradish peroxidase (HRP) was found to be a well-suited reagent for the detection of Ag-bound scFvs in comparison with either anti-c-myc Ab or the mixing procedure. Based on our findings, it seems there is no universal media supplement for an improved expression of all scFvs derived from both Tomlinson I and J libraries. We thus propose that expression of scFv fragments in a microplate scale is largely dependent on a variety of parameters, in particular the scFv clones and relevant sequences.

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Phage Display on the Base of Filamentous Bacteriophages: Application for Recombinant Antibodies Selection

Cited by 36 publications

References 85 publications

Phage display of functional αβ single-chain T-cell receptor molecules specific for CD1b:Ac2SGL complexes from Mycobacterium tuberculosis-infected cells

Phage display of functional αβ single-chain T-cell receptor molecules specific for CD1b:Ac2SGL complexes from Mycobacterium tuberculosis-infected cells

Architectural Insight into Inovirus-Associated Vectors (IAVs) and Development of IAV-Based Vaccines Inducing Humoral and Cellular Responses: Implications in HIV-1 Vaccines

Improved Soluble ScFv ELISA Screening Approach for Antibody Discovery Using Phage Display Technology

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