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phage display

Yin-Ku Liang, Liu-Jiao Bian
Kringle 5, the fifth fragment of plasminogen, is known to be important for inhibiting the proliferation and migration of vascular endothelial cell (VEC), while not having any effects on normal endothelial cells. Therefore, it may be a potential tumor therapy candidate. However, the ligand of the Kringle 5 in VEC has not yet been identified. In this study, the possible ligand of Kringle 5 in vitro was screened and validated using Ph.D.-7 phage display peptide library with molecular docking, along with surface plasma resonance (SPR)...
2016: PloS One
Yubao Cui, Lili Yu, Ying Zhou, Li Yang, Chengbo Zhang
Mimotope mapping enables the characterization of allergen epitopes for the development of diagnostic and therapeutic approaches. In the present study, a phage display peptide library was used for mimotope mapping based on the binding of antibodies against the recombinant group 5 allergen from the house dust mite Dermatophagoides farinae (Der f 5), an arthropod that causes indoor allergies worldwide. When three monoclonal anti‑Der f 5 antibodies were used for biopanning, seven mimotopes were identified. Their common subsequence was '‑‑‑[‑A][‑T]W[‑S]H[HSFW][LM][PSKR] [TLV][AST]‑[DP][‑L]‑'...
October 13, 2016: Molecular Medicine Reports
Katarzyna Szot-Karpińska, Piotr Golec, Adam Lesniewski, Barbara Palys, Frank Marken, Joanna Niedziółka-Jönsson, Grzegorz Wegrzyn, Marcin Łoś
With the advent of nanotechnology, carbon nanomaterials such as carbon nanofibers (CNF) have aroused substantial interest in various research fields including energy storage and sensing. Further improvement of their properties might be achieved via application of viral particles such as bacteriophages. In this report we present a filamentous M13 bacteriophage, with a point mutation in gene VII (pVII-mutant-M13), that selectively binds to the carbon nanofibers to form 3D structures. The phage-display technique was utilized for the selection of the pVII-mutant-M13 phage from the phage display peptide library...
October 17, 2016: Bioconjugate Chemistry
Ricardo Jara-Acevedo, Paula Díez, María González-González, Rosa María Dégano, Nieves Ibarrola, Rafael Góngora, Alberto Orfao, Manuel Fuentes
The selection process aims sequential enrichment of phage antibody display library in clones that recognize the target of interest or antigen as the library undergoes successive rounds of selection. In this review, selection methods most commonly used for phage display antibody libraries have been comprehensively described.
October 7, 2016: Current Pharmaceutical Design
Kristin Ciezki, Kristen Murfin, Heidi Goodrich-Blair, S Patricia Stock, Steven Forst
R-type bacteriocins are contractile phage-tail-like structures that are bactericidal towards related bacterial species. The C-terminal region of the phage tail fiber protein determines target-binding specificity. The mutualistic bacteria Xenorhabdus nematophila and Xenorhabdus bovienii produce R-type bacteriocins (xenorhabdicins) that are selectively active against different Xenorhabdus species. We analyzed the P2-type remnant prophage clusters in draft sequences of nine strains of X. bovienii The C-terminal tail fiber region in each of the respective strains was unique and consisted of mosaics of modular units...
October 12, 2016: FEMS Microbiology Letters
Stefano Mancini, Carmen Menzi, Frank Oechslin, Philippe Moreillon, José Manuel Entenza
Streptococcus gordonii and related species of oral viridans group streptococci (VGS) are common etiological agents of infective endocarditis (IE). We explored vaccination as a strategy to prevent VGS- IE, using a novel antigen presenting system based on non-genetically modified Lactococcus lactis displaying vaccinogens on its surface.Hsa and PadA are surface-located S. gordonii proteins implicated in platelet adhesion and aggregation, which are key steps in the pathogenesis of IE. This function makes them ideal targets for vaccination against VGS- IE...
October 10, 2016: Infection and Immunity
Silvia Sonzini, Alessio Marcozzi, Raphael J Gubeli, Christopher F van der Walle, Peter Ravn, Andreas Herrmann, Oren A Scherman
Supramolecular interactions between the host cucurbit[8]uril (CB[8]) and amino acids have been widely interrogated, but recognition of specific motifs within a protein domain have never been reported. A phage display approach was herein used to select motifs with the highest binding affinity for the heteroternary complex with methyl viologen and CB[8] (MV⋅CB[8]) within a vast pool of cyclic peptide sequences. From the selected motifs, an epitope consisting of three amino acid was extrapolated and incorporated into a solvent-exposed loop of a protein domain; the protein exhibited micromolar binding affinity for the MV⋅CB[8] complex, matching that of the cyclic peptide...
October 13, 2016: Angewandte Chemie
M Brasino, J N Cha
Here we describe novel covalent conjugates of antibody-phage for the detection of multiple cancer biomarkers using real time immuno-polymerase chain reaction (immuno-PCR). While the conventional process of immuno-PCR utilizes DNA-conjugated antibodies, chemical modification of antibodies not only reduces antibody affinity but also creates a heterogeneous population of products. However, phage naturally encapsulate genomic DNA, which can be used as a PCR template. To produce covalently conjugated antibody-phage constructs without recombinant antibody expression or chemical modification of antibodies, we incorporated a photocrosslinkable non-canonical amino acid within an antibody-binding domain displayed on one of the phage coat proteins...
October 12, 2016: Analyst
William J J Finlay, Laird Bloom, Sreeja Varghese, Bénédicte Autin, Orla Cunningham
High-affinity, highly specific binding proteins are a key class of molecules used in the development of new affinity chromatography methods. Traditionally, antibody-based methods have relied on the use of immunoglobulins purified from immune animal sera, from egg yolks, or from murine monoclonal hybridoma supernatants. To accelerate and refine the reagent antibody generation process, we have developed optimized methods that allow the rapid assembly of scFv libraries from chickens immunized with pools of immunogens...
2017: Methods in Molecular Biology
William J J Finlay, Laird Bloom, Joanne Grant, Edward Franklin, Deirdre Ní Shúilleabháin, Orla Cunningham
Antibodies are critical reagents in many fundamental biochemical methods such as affinity chromatography, enzyme-linked immunosorbent assays (ELISA), flow cytometry, western blotting, immunoprecipitation, and immunohistochemistry techniques. As our understanding of the proteome becomes more complex, demand is rising for rapidly generated antibodies of higher specificity than ever before. It is therefore surprising that few investigators have moved beyond the classical methods of antibody production in their search for new reagents...
2017: Methods in Molecular Biology
Carmen Burtea, Sophie Laurent, Tuba Sanli, Deborah Fanfone, Aude Devalckeneer, Sébastien Sauvage, Marie-Claire Beckers, Sandrine Rorive, Isabelle Salmon, Luce Vander Elst, Bernard R Lauwerys, Robert N Muller
BACKGROUND: Interleukin-7 receptor alpha (IL-7Rα) represents a biomarker with potential applications in rheumatoid arthritis (RA) diagnosis and therapy. We have therefore searched by phage display potential IL-7Rα specific peptides with the primary goal being to develop in vivo molecular imaging tools. METHODS: IL-7Rα-targeted peptides were searched within a disulfide-constrained combinatorial phage displayed library of random linear heptapeptides. The apparent dissociation constant (Kd) and half maximal inhibition constant (IC50) were estimated for phage clones and synthesized peptides by ELISA...
October 12, 2016: Arthritis Research & Therapy
Jun-Haeng Cho, A-Ru Kim, Sang-Heon Kim, Su-Jae Lee, Hoeil Chung, Moon-Young Yoon
CD133 is known as biomarker for glioblastoma (GBM) and also serves as a marker for cancer stem cells (CSCs), which carry out tumorigenesis and resist conventional therapeutics. The presence of CD133-presenting CSC is a one of the factors in maintenance of the tumorigenic potential of GBM. Thus, CD133 is a potential target for accurate diagnosis of GBM, which could improve its poor prognosis for patients when CSCs are present. Herein we designed a small peptide-based imaging agent with stimulus-responsive properties...
October 6, 2016: Acta Biomaterialia
Ali Akbar Alizadeh, Maryam Hamzeh-Mivehroud, Malak Farajzadeh, Siavoush Dastmalchi
The aim of this study was to identify novel TNF-α blocking peptide(s) using phage display technology. Two novel 7-mer TNF-α binding peptides P51 and P52 with Kd values of 1.47 and 0.51nM were identified. Phage particles displaying P51 and P52 peptides at 0.318nM concentration prevent cytotoxic effect of TNF-α on L929 cells by 8.2% and 16.15%, respectively. Synthesized P51 and P52 peptides also inhibited TNF-α induced cytotoxicity with IC50 values of 25.15±2.18 and 7.08±2.24μM, respectively. The result of RT-PCR also supports the inhibitory activity of the identified peptides, where P51 and P52 significantly inhibit the inductive effect of TNF-α on IκB-α mRNA levels...
October 5, 2016: European Journal of Pharmaceutical Sciences
Gene Kurosawa, Mariko Kondo, Yoshikazu Kurosawa
When the technology for constructing human antibody (Ab) libraries using a phage-display system was developed, many researchers in Ab-related fields anticipated that it would be widely applied to the development of pharmaceutical drugs against various diseases, including cancers. However, successful examples of such applications are very limited. Moreover, researchers who utilize phage-display technology now show divergent ways of thinking about phage Ab libraries. For example, there is debate about what should be the source of VH and VL genes for the construction of libraries to cover the whole repertoire of Abs present in the human body...
October 5, 2016: Biochemical and Biophysical Research Communications
Kotaro Sakamoto, Yayoi Kawata, Yasushi Masuda, Tadashi Umemoto, Takashi Ito, Taiji Asami, Shiro Takekawa, Tetsuya Ohtaki, Hiroshi Inooka
Fibroblast growth factor receptor-1c (FGFR1c)/βKlotho (KLB) complex is a receptor of fibroblast growth factor 21 (FGF21). Pharmacologically, FGF21 shows anti-obesity and anti-diabetic effects upon peripheral administration. Here, we report the development of an artificial peptide agonist to the FGFR1c/KLB heterodimer complex. The peptide, F91-8A07 (LPGRTCREYPDLWWVRCY), was discovered from random peptide T7 phage display and selectively bound to the FGFR1c/KLB complex, but not to FGFR1c and KLB individually...
October 4, 2016: Biochemical and Biophysical Research Communications
Jan Müller, Robin Reichel, Sebastian Vogt, Stefan P Müller, Wolfgang Sauerwein, Wolfgang Brandau, Angelika Eggert, Alexander Schramm
Neuroectodermal tumours are characterized by aberrant processing of disialogangliosides concomitant with high expression of GD2 or GD3 on cell surfaces. Antibodies targeting GD2 are already in clinical use for therapy of neuroblastoma, a solid tumour of early childhood. Here, we set out to identify peptides with high affinity to human disialoganglioside GD2. To this end, we performed a combined in vivo and in vitro screen using a recombinant phage displayed peptide library. We isolated a phage displaying the peptide sequence WHWRLPS that specifically binds to the human disialoganglioside GD2...
2016: PloS One
Wei Li, Hongjia Yang, Dimiter S Dimitrov
CD16A (FcγRIIIA) is an activating receptor mostly expressed on natural killer (NK) cells and monocytes/macrophages. It can mediate antibody-dependent cell-mediated cytotoxicity (ADCC) through low-affinity interaction with human immunoglobulin G (IgG) Fc. It can also mediate cell lysis if NK cells are guided by bispecific killer cells engagers (BiKEs). BiKEs showed some success in clinical trials of cancer and are promising candidate therapeutics. However, currently reported BiKEs are based on antibody fragments (scFvs) of relatively large size...
October 3, 2016: Experimental and Molecular Pathology
George Kubas, William Rees, Jonathan Caguiat, David Asch, Diana Fagan, Pedro Cortes
The present research investigates the identification of amino acid sequences that selectively bind to a pentaerythritol tetranitrate (PETN) explosive surrogate. Through the use of a phage display technique and enzyme-linked immunosorbent assays (ELISA), a peptide library was tested against PETNH (pentaerythritol trinitrate hemisuccinate), a surrogate of PETN, in order to screen for those with amino acids having affinity toward the explosive. The use of the PETN surrogate allowed the immobilization of the target during the screening process, while retaining a chemical profile similar to that of PETN...
October 6, 2016: Biopolymers
Jonas Zantow, Sarah Just, Ilias Lagkouvardos, Sigrid Kisling, Stefan Dübel, Patricia Lepage, Thomas Clavel, Michael Hust
Pathogen infections, autoimmune diseases, and chronic inflammatory disorders are associated with systemic antibody responses from the host immune system. Disease-specific antibodies can be important serum biomarkers, but the identification of antigens associated with specific immune reactions is challenging, in particular if complex communities of microorganisms are involved in the disease progression. Despite promising new diagnostic opportunities, the discovery of these serological markers becomes more difficult with increasing complexity of microbial communities...
October 5, 2016: Scientific Reports
Sally Elizabeth Hall, Brett Nixon, R John Aitken
Feral horses are a significant pest species in many parts of the world, contributing to land erosion, weed dispersal and the loss of native flora and fauna. There is an urgent need to modify feral horse management strategies to achieve public acceptance and long-term population control. One way to achieve this is by using non-surgical methods of sterilisation, which are suitable in the context of this mobile and long-lived species. In this review we consider the benefits of implementing novel mechanisms designed to elicit a state of permanent sterility (including redox cycling to generate oxidative stress in the gonad, random peptide phage display to target non-renewable germ cells and the generation of autoantibodies against proteins essential for conception via covalent modification) compared with that of traditional immunocontraceptive approaches...
October 5, 2016: Reproduction, Fertility, and Development
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