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https://www.readbyqxmd.com/read/28526816/delineation-of-b-cell-epitopes-of-salmonella-enterica-serovar-typhi-hemolysin-e-potential-antibody-therapeutic-target
#1
Chai Fung Chin, Jing Yi Lai, Yee Siew Choong, Amy Amilda Anthony, Asma Ismail, Theam Soon Lim
Hemolysin E (HlyE) is an immunogenic novel pore-forming toxin involved in the pathogenesis of typhoid fever. Thus, mapping of B-cell epitopes of Salmonella enterica serovar Typhi (S. Typhi) is critical to identify key immunogenic regions of HlyE. A random 20-mer peptide library was used for biopanning with enriched anti-HlyE polyclonal antibodies from typhoid patient sera. Bioinformatic tools were used to refine, analyze and map the enriched peptide sequences against the protein to identify the epitopes. The analysis identified both linear and conformational epitopes on the HlyE protein...
May 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28513737/dimeric-and-trimeric-derivatives-of-the-azinomycin-b-chromophore-show-enhanced-dna-binding
#2
Milena Balazy, Alejandra Fausto, Christina Voskanian, Bianca Chavez, Harmanpreet Panesar, Thomas G Minehan
To explore the utility of the azinomycin B chromophore as a platform for the development of major-groove binding small molecules, we have prepared a series of 3-methoxy-5-methylnaphthalene derivatives containing diamine, triamine, and carbohydrate linker moieties. All bis- and tris-azinomycin derivatives are intercalators that display submicromolar binding affinities for calf-thymus DNA, as revealed by viscometry measurements and fluorescent intercalator displacement (FID) assays, respectively. Although the tightest binding ligand 1d (Ka = 2...
May 17, 2017: Organic & Biomolecular Chemistry
https://www.readbyqxmd.com/read/28503119/targeting-hiv-1-envelope-proteins-using-a-fragment-discovery-all-atom-computational-algorithm
#3
Michael H Peters
INTRODUCTION: HIV viral envelope proteins are targets for small inhibitor molecules aimed at disrupting the cellular entry process. Potential peptide-class inhibitor molecules (rDNA drugs) have been previously identified, with mixed results, through biomimicry and phage display experimental methods. Here we describe a new approach based on computational fragment discovery. The method has the potential to not only optimize peptide binding affinity but also to rapidly produce alternative inhibitors against mutated strains...
April 2017: Current Enzyme Inhibition
https://www.readbyqxmd.com/read/28498803/differentially-expressed-proteins-in-glioblastoma-multiforme-identified-with-a-nanobody-based-anti-proteome-approach-and-confirmed-by-oncofinder-as-possible-tumor-class-predictive-biomarker-candidates
#4
Ivana Jovčevska, Neja Zupanec, Žiga Urlep, Andrej Vranič, Boštjan Matos, Clara Limbaeck Stokin, Serge Muyldermans, Michael P Myers, Anton A Buzdin, Ivan Petrov, Radovan Komel
Glioblastoma multiforme is the most frequent primary malignancy of the central nervous system. Despite remarkable progress towards an understanding of tumor biology, there is no efficient treatment and patient outcome remains poor. Here, we present a unique anti-proteomic approach for selection of nanobodies specific for overexpressed glioblastoma proteins. A phage-displayed nanobody library was enriched in protein extracts from NCH644 and NCH421K glioblastoma cell lines. Differential ELISA screenings revealed seven nanobodies that target the following antigens: the ACTB/NUCL complex, VIM, NAP1L1, TUFM, DPYSL2, CRMP1, and ALYREF...
April 24, 2017: Oncotarget
https://www.readbyqxmd.com/read/28498372/comparative-transcriptomics-analyses-reveal-the-conservation-of-an-ancestral-infectious-strategy-in-two-bacteriophage-genera
#5
Bob G Blasdel, Anne Chevallereau, Marc Monot, Rob Lavigne, Laurent Debarbieux
Although the evolution of tailed bacteriophages has increasingly been better understood through comparisons of their DNA sequences, the functional consequences of this evolution on phage infectious strategies have remained unresolved. In this study, we comprehensively compared the transcriptional strategies of two related myoviruses, PAK_P3 and PAK_P4, infecting the same Pseudomonas aeruginosa host strain. Outside of the conservation of their structural clusters, their highly syntenic genomes display only limited DNA similarity...
May 12, 2017: ISME Journal
https://www.readbyqxmd.com/read/28494872/using-resurrected-ancestral-proviral-proteins-to-engineer-virus-resistance
#6
Asunción Delgado, Rocio Arco, Beatriz Ibarra-Molero, Jose M Sanchez-Ruiz
Proviral factors are host proteins hijacked by viruses for processes essential for virus propagation such as cellular entry and replication. Pathogens and their hosts co-evolve. It follows that replacing a proviral factor with a functional ancestral form of the same protein could prevent viral propagation without fatally compromising organismal fitness. Here, we provide proof of concept of this notion. Thioredoxins serve as general oxidoreductases in all known cells. We report that several laboratory resurrections of Precambrian thioredoxins display substantial levels of functionality within Escherichia coli...
May 9, 2017: Cell Reports
https://www.readbyqxmd.com/read/28494149/comparison-of-three-anti-hapten-vhh-selection-strategies-for-the-development-of-highly-sensitive-immunoassays-for-microcystins
#7
Macarena Pírez-Schirmer, Martin A Rossotti, Natalia Badagian, Carmen Leizagoyen, Beatriz Margarita Brena, Gualberto G Gonzalez-Sapienza
Owing to their reproducibility, stability and cost-effective production, the recombinant variable domains of heavy-chain-only antibodies (VHHs) are becoming a salient option as immunoassay reagents. Recently, there have been several reports describing their application to the detection of small molecules (haptens). However, lacking the heavy-light chain interface of conventional antibodies, VHHs are not particularly apt to bind small analytes and failures are not uncommon. Here we describe the construction of a VHH phage display library against the cyanobacterial hepatotoxin microcystin LR, and its selection using competitive panning and two novel panning strategies...
May 11, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28491282/cell-growth-inhibition-and-apoptotic-effects-of-a-specific-anti-rtfscfv-antibody-on-prostate-cancer-but-not-glioblastoma-cells
#8
Foroogh Nejatollahi, Payam Bayat, Bahareh Moazen
Background: Single chain antibody (scFv) has shown interesting results in cancer immunotargeting approaches, due to its advantages over monoclonal antibodies. Regeneration and tolerance factor (RTF) is one of the most important regulators of extracellular and intracellular pH in eukaryotic cells. In this study, the inhibitory effects of a specific anti-RTF scFv were investigated and compared between three types of prostate cancer and two types of glioblastoma cells.  Methods: A phage antibody display library of scFv was used to select specific scFvs against RTF using panning process...
2017: F1000Research
https://www.readbyqxmd.com/read/28488699/charting-the-unknown-epitranscriptome
#9
Eva Maria Novoa, Christopher E Mason, John S Mattick
RNA modifications can alter RNA structure-function relationships and various cellular processes. However, the genomic distribution and biological roles of most RNA modifications remain uncharacterized. Here, we propose using phage display antibody technology and direct sequencing through nanopores to facilitate systematic interrogation of the distribution, location and dynamics of RNA modifications.
May 10, 2017: Nature Reviews. Molecular Cell Biology
https://www.readbyqxmd.com/read/28484722/bacteriophages-and-their-immunological-applications-against-infectious-threats
#10
REVIEW
Elena Criscuolo, Sara Spadini, Jacopo Lamanna, Mattia Ferro, Roberto Burioni
Bacteriophage therapy dates back almost a century, but the discovery of antibiotics led to a rapid decline in the interests and investments within this field of research. Recently, the novel threat of multidrug-resistant bacteria highlighted the alarming drop in research and development of new antibiotics: 16 molecules were discovered during 1983-87, 10 new therapeutics during the nineties, and only 5 between 2003 and 2007. Phages are therefore being reconsidered as alternative therapeutics. Phage display technique has proved to be extremely promising for the identification of effective antibodies directed against pathogens, as well as for vaccine development...
2017: Journal of Immunology Research
https://www.readbyqxmd.com/read/28483948/high-affinity-internalizing-human-scfv-fc-antibody-for-targeting-fgfr1-overexpressing-lung-cancer
#11
Aleksandra Sokolowska-Wedzina, Grzegorz Chodaczek, Julia Chudzian, Aleksandra Borek, Malgorzata Zakrzewska, Jacek Otlewski
Targeted delivery of anti-cancer drugs using antibodies specific for tumor-associated antigens represents one of the most important approaches in current immuno-oncology research. Fibroblast growth factor receptor 1 (FGFR1) has been demonstrated to be a high-frequency targetable oncogene specific for smoking-associated lung cancers, present in over 20% of lung squamous cell carcinoma cases. This report describes the generation of a potent, fully human antibody fragment in scFv-Fc format efficiently targeting FGFR1...
May 8, 2017: Molecular Cancer Research: MCR
https://www.readbyqxmd.com/read/28482148/bacteriophages-as-factories-for-eu2o3-nanoparticle-synthesis
#12
Piotr Golec, Kamila Żelechowska, Joanna Karczewska-Golec, Jakub Karczewski, Adam Lesniewski, Marcin Łoś, Grzegorz Wegrzyn, Andrzej M Klonkowski
The use of phage display to identify peptides with an ability to bind Eu2O3 is demonstrated in this report. This is the first report of modified phages specifically binding a lanthanide. The peptides exposed on virions revealed very strong binding to Eu2O3 nanoparticles and the ability to catalyze Eu2O3 nanoparticles' formation from Eu(OH)3 and Eu(NO3)3 solutions. The luminescence emission spectrum of Eu3+ ions indicated that these ions existed mostly in sites deviated from the inversion symmetry in crystalline Eu2O3 aggregates and gelatinous Eu(OH)3 precipitate...
May 8, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28475831/microarray-based-immunoassay-with-synthetic-mimotopes-for-the-detection-of-fumonisin-b1
#13
Riikka Peltomaa, Elena Benito-Peña, Rodrigo Barderas, Ursula Sauer, Martin González-Andrade, Maria C Moreno-Bondi
Mimotopes, or epitope mimics, can be applied to competitive immunoassays, for the detection of low molecular weight natural toxicants, as an alternative to toxin-conjugates. In this work, we report the development of a microarray-based immunoassay for the detection of fumonisin B1 using a novel mimotope selected by phage display technology. Fumonisin-specific antibody was used to isolate mimotopes from a 12-mer peptide library in successive selection rounds. Enrichment of antibody binding phages was observed after three panning rounds, and sequence analysis of randomly selected monoclonal phages revealed two conserved peptide sequences...
May 5, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28475681/a-camel-anti-lysozyme-cdr3-only-domain-antibody-selected-from-phage-display-vhh-library-acts-as-potent-lysozyme-inhibitor
#14
Lingling Qiu, Yaning Feng, Xiaoling Ma, Jiangwei Li
Mimetics of antibody-binding sites represent particularly interesting targets, however they are difficult to identify. In most cases, naturally derived CDR3 peptides show a much lower activity and affinity. In this study, we identified a CDR3 domain antibody with framework 3 (FR3) and FR4 in the flank by screening a lysozyme-immunized phage display VHH library. This antibody has a potent enzyme inhibiting activity and high thermal stability. With sequence alignment and site-directed mutagenic analysis, we found that the cysteine residue at amino acid position 88 in FR3 might play a key role in maintaining the stability of the CDR3 antibody...
May 5, 2017: Acta Biochimica et Biophysica Sinica
https://www.readbyqxmd.com/read/28472478/improved-antibody-based-ricin-neutralization-by-affinity-maturation-is-correlated-with-slower-off-rate-values
#15
Ronit Rosenfeld, Ron Alcalay, Adva Mechaly, Gideon Lapidoth, Eyal Epstein, Chanoch Kronman, Sarel J Fleishman, Ohad Mazor
While potent monoclonal antibodies against ricin were introduced over the years, the question whether increasing antibody affinity enables better toxin neutralization was not fully addressed yet. The aim of this study was to characterize the contribution of antibody affinity to the ricin neutralization potential of the antibody. cHD23 monoclonal antibody that targets the toxin B-subunit and interferes with its binding to membranal receptors, was isolated. In order to create antibody clones with improved affinity toward ricin, a scFv-phage display library containing mutated versions of the variable regions of cHD23 was constructed and clones with improved binding of ricin were isolated...
May 3, 2017: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/28470616/in-vivo-biotinylation-of-antigens-in-e-coli
#16
Susanne Gräslund, Pavel Savitsky, Susanne Müller-Knapp
Site-specific biotinylation of proteins is often the method of choice to enable efficient immobilization of a protein on a surface without interfering with protein folding. The tight interaction of biotin and streptavidin is frequently used to immobilize an antigen during phage display selections of binders. Here we describe a method of in vivo biotinylation of proteins during expression in E. coli, by tagging the protein with the short biotin acceptor peptide sequence, Avi tag, and co-expression of the E. coli biotin ligase (BirA) resulting in precise biotinylation of a specific lysine residue in the tag...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28469983/two-epitopes-responsible-for-the-catalytic-activity-of-heme-oxygenase-1-identified-by-phage-display
#17
Xuran Wei, Qingjun Liu, Yaping Gao, Jun Yang, Bo Wang, Guang Yang, Shihui Zhang, Hong Zhou
Heme oxygenase-1 (HO-1) catalyzes the oxidative degradation of heme. The catalytic mechanism of the HO-1 reaction has been determined gradually by studies of its crystal structure and HO-1 mutants. However, the neutralizing epitopes responsible for HO-1 activity remain elusive. Screening of a phage display library revealed four epitopes that could interact with the polyclonal antibody prepared by immunizing rabbits with the purified HO-1 protein. Two of these four epitopes are responsible for HO-1 catalytic activity because their antibodies were able to neutralize HO-1 activity...
May 2017: FEBS Open Bio
https://www.readbyqxmd.com/read/28468949/identification-of-a-novel-the-itg%C3%AE-v%C3%AE-6-binding-peptide-using-protein-separation-and-phage-display
#18
Uwe Haberkorn, Annette Altmann, Max Sauter, Saskia Roesch, Walter Mier, Rolf Warta, Juergen Debus, Gerhard Dyckhoff, Christel Herold-Mende
PURPOSE: Targeted therapies are regarded as promising approaches to increase 5-year survival rate of head and neck squamous cell carcinoma (HNSCC) patients. EXPERIMENTAL DESIGN: For the selection of carcinoma-specific peptides membrane proteome of HNO97 tumor cells fractionated by the ProteomeLab™ PF2D system and corresponding HNO97 cells were deployed for an alternating biopanning using a sunflower trypsin inhibitor1-based phage display (SFTI8Ph) library. Stability, binding properties and affinity of novel candidates were assessed in vitro using radio-HPLC, binding experiments and surface plasmon resonance assay (SPR), respectively...
May 3, 2017: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28468775/targeting-insulin-receptor-in-breast-cancer-using-small-engineered-protein-scaffolds
#19
Jie Ying Chan, Benjamin J Hackel, Douglas Yee
Insulin receptor (InsR) and the type I insulin-like growth factor (IGF1R) are homologous receptors necessary for signal transduction by their cognate ligands insulin, insulin-like growth factor-I and -II (IGF-I and IGF-II). IGF1R monoclonal antibodies, intended to inhibit malignant phenotypic signaling, failed to show benefit in patients with endocrine-resistant tumors in phase III clinical trials. Our previous work showed that in tamoxifen-resistant cells IGF1R expression was lacking but InsR inhibition effectively blocked growth...
May 3, 2017: Molecular Cancer Therapeutics
https://www.readbyqxmd.com/read/28468593/hyaluronan-binding-peptide-for-targeting-peritoneal-carcinomatosis
#20
Hideki Ikemoto, Prakash Lingasamy, Anne-Mari Anton Willmore, Hedi Hunt, Kaarel Kurm, Olav Tammik, Pablo Scodeller, Lorena Simón-Gracia, Venkata Ramana Kotamraju, Andrew M Lowy, Kazuki N Sugahara, Tambet Teesalu
Peritoneal carcinomatosis results from dissemination of solid tumors in the peritoneal cavity, and is a common site of metastasis in patients with carcinomas of gastrointestinal or gynecological origin. Peritoneal carcinomatosis treatment is challenging as poorly vascularized, disseminated peritoneal micro-tumors are shielded from systemic anticancer drugs and drive tumor regrowth. Here, we describe the identification and validation of a tumor homing peptide CKRDLSRRC (IP3), which upon intraperitoneal administration delivers payloads to peritoneal metastases...
May 2017: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
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