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phage display

Cintia P J Rua, Louisi S de Oliveira, Adriana Froes, Diogo A Tschoeke, Ana Carolina Soares, Luciana Leomil, Gustavo B Gregoracci, Ricardo Coutinho, Eduardo Hajdu, Cristiane C Thompson, Roberto G S Berlinck, Fabiano L Thompson
Marine sponge holobionts harbor complex microbial communities whose members may be the true producers of secondary metabolites accumulated by sponges. Bromopyrrole alkaloids constitute a typical class of secondary metabolites isolated from sponges that very often display biological activities. Bromine incorporation into secondary metabolites can be catalyzed by either halogenases or haloperoxidases. The diversity of the metagenomes of sponge holobiont species containing bromopyrrole alkaloids (Agelas spp. and Tedania brasiliensis) as well as holobionts devoid of bromopyrrole alkaloids spanning in a vast biogeographic region (approx...
March 15, 2018: Microbial Ecology
Seung Yub Han, Alesia Antoine, David Howard, Bryant Chang, Woo Sung Chang, Matthew Slein, Gintaras Deikus, Sofia Kossida, Patrice Duroux, Marie-Paule Lefranc, Robert P Sebra, Melissa L Smith, Ismael Ben F Fofana
The simian immunodeficiency virus (SIV)/macaque model of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome pathogenesis is critical for furthering our understanding of the role of antibody responses in the prevention of HIV infection, and will only increase in importance as macaque immunoglobulin (IG) gene databases are expanded. We have previously reported the construction of a phage display library from a SIV-infected rhesus macaque ( Macaca mulatta ) using oligonucleotide primers based on human IG gene sequences...
2018: Frontiers in Immunology
V Vasilca, A Sadeghpour, S Rawson, L E Hawke, S A Baldwin, T Wilkinson, D Bannister, V L G Postis, M Rappolt, S P Muench, L J C Jeuken
Screening assays performed against membrane protein targets (e.g. phage display) are hampered by issues arising from protein expression and purification, protein stability in detergent solutions and epitope concealment by detergent micelles. Here, we have studied a fast and simple method to improve screening against membrane proteins: spherical-supported bilayer lipid membranes ("SSBLM"). SSBLMs can be quickly isolated via low-speed centrifugation and redispersed in liquid solutions while presenting the target protein in a native-like lipid environment...
March 12, 2018: Analytical Biochemistry
Fatemeh Faraji, Nader Tajik, Mahdi Behdani, Mohammad Ali Shokrgozar, Amir Hassan Zarnani, Fatemeh Shahhosseini, Mahdi Habibi-Anbouhi
CD22 is a B-cell-specific trans-membrane glycoprotein which is found on the surface of the most B-cells and modulates their function, survival, and apoptosis. Recently, targeting this cell surface biomarker in B-cell malignancies and disorders has attracted a lot of attention. The variable domain of camelid single chain antibodies (VHH, Nanobody) is a form of antibodies with novel properties including small size (15-17 kDa), thermal and chemical stability, high affinity and homology to human antibody sequences...
March 15, 2018: Biotechnology and Applied Biochemistry
Pooja Sadana, Rebecca Geyer, Joern Pezoldt, Saskia Helmsing, Jochen Huehn, Michael Hust, Petra Dersch, Andrea Scrima
Yersinia pseudotuberculosis is a Gram-negative bacterium and zoonotic pathogen responsible for a wide range of diseases, ranging from mild diarrhea, enterocolitis, lymphatic adenitis to persistent local inflammation. The Y. pseudotuberculosis invasin D (InvD) molecule belongs to the invasin (InvA)-type autotransporter proteins, but its structure and function remain unknown. In this study, we present the first crystal structure of InvD, analyzed its expression and function in a murine infection model, and identified its target molecule in the host...
March 13, 2018: Journal of Biological Chemistry
André Schiefner, Michaela Gebauer, Antonia Richter, Arne Skerra
We describe the comparative X-ray structural analysis of three Anticalin proteins directed against the extra-domain B (ED-B) of oncofetal fibronectin (Fn), a validated marker of tumor neoangiogenesis. The Anticalins were engineered from the human lipocalin 2 (Lcn2) scaffold via targeted randomization of the structurally variable loop region and selection by phage display, resulting in 15-19 exchanged residues. While the four reshaped loops exhibit diverse conformations (with shifts in Cα positions up to 20...
March 5, 2018: Structure
Abbas Shali, Sadegh Hasannia, Fatemeh Gashtasbi, S Masoud Abdous, S Shirin Shahangian, Shirin Jalili
Since anthrax is an acute infectious disease, detection and neutralization of the toxins of pathogenic Bacillus anthracis are of great importance. The critical role of protective antigen (PA) component of tripartite anthrax toxin in toxin entry into the host cell cytosol provided a great deal of effort to generate monoclonal antibodies against this constitute. Regarding the importance of anthrax detection/neutralization and unique physicochemical and pharmacological features of VHHs as single domain antibodies, the present study aimed to generate VHHs against the receptor binding domain of PA, termed PAD4...
March 7, 2018: International Journal of Biological Macromolecules
Mengyao Wang, Yuhua Wei, Wei Yu, Lizi Wang, Lu Zhai, Xiaoting Li, Xintong Wang, Hua Zhang, Zhenyue Feng, Liquan Yu, Yongzhong Yu, Jinzhu Ma, Yudong Cui
The GapC protein of Staphylococcus aureus (S. aureus) is a surface protein that is highly conserved among Staphylococcus strains, and it can induce protective humoral immune responses. However, B-cell epitopes in S. aureus GapC have not been reported. In this study, we generated a monoclonal antibody (mAb2A9) targeting S. aureus GapC. Through a passive immunity test, mAb2A9 was shown to partially protect mice against S. aureus infection. We screened the motif236 PVATGSLTE243 that is recognized by mAb2A9 using a phage-display system...
March 6, 2018: Microbial Pathogenesis
Jiao Wang, Jingjing Song, Shuimei Zhou, Yourong Fu, Jeffrey A Bailey, Changxin Shen
BACKGROUND: Identification of RhD antigen epitopes is a key component in understanding the pathogenesis of haemolytic disease of the foetus and newborn. Research has indicated that phage display libraries are useful tools for identifying novel mimic epitopes (mimotopes) which may help to determine antigen specificity. MATERIALS AND METHODS: We selected the mimotopes of blood group RhD antigen by affinity panning a phage display library using monoclonal anti-D. After three rounds of biopanning, positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA) and then sent for sequencing and peptides synthesis...
January 16, 2018: Blood Transfusion, Trasfusione del Sangue
Daniel P Teufel, Gavin Bennett, Helen Harrison, Katerine van Rietschoten, Silvia Pavan, Catherine Stace, François Le Floch, Tine Van Bergen, Elke Vermassen, Philippe Barbeaux, Tjing-Tjing Hu, Jean H M Feyen, Marc Vanhove
Plasma kallikrein, a member of the kallikrein-kinin system, catalyzes the release of the bioactive peptide bradykinin, which induces inflammation, vasodilation, vessel permeability and pain. Preclinical evidence implicates the activity of plasma kallikrein in diabetic retinopathy, which is a leading cause of visual loss in patients suffering from diabetes mellitus. Employing a technology based on phage-display combined with chemical cyclization, we have identified highly selective bicyclic peptide inhibitors with nano- and pico-molar potencies towards plasma kallikrein...
March 8, 2018: Journal of Medicinal Chemistry
Claudine Boiziau, Macha Nikolski, Elodie Mordelet, Justine Aussudre, Karina Vargas-Sanchez, Klaus G Petry
Multiple sclerosis is characterized by inflammatory lesions dispersed throughout the central nervous system (CNS) leading to severe neurological handicap. Demyelination, axonal damage, and blood brain barrier alterations are hallmarks of this pathology, whose precise processes are not fully understood. In the experimental autoimmune encephalomyelitis (EAE) rat model that mimics many features of human multiple sclerosis, the phage display strategy was applied to select peptide ligands targeting inflammatory sites in CNS...
March 7, 2018: Inflammation
Eun Sook Kim, Hee-Won Bae, You-Hee Cho
The host range of a phage is determined primarily by phage-receptor interaction. Here, we profiled the host range of an RNA leviphage, PP7 that requires functional type IV pilus (TFP) in order to enter into its host bacterium, Pseudomonas aeruginosa . Out of 25 twitching-proficient P. aeruginosa strains, 4 with group I pilin and 7 with group III pilin displayed PP7-resistance. The remaining 14 possessed group II pilin, which included 10 PP7-sensitive and 4 PP7-resistant strains, suggesting that only the strains with TFP consisted of a subset of group II (hence, group IIa) pilin were susceptible to PP7...
2018: Frontiers in Microbiology
Federico M Winkler, Ricardo García, María V Valdivia, Karin B Lohrmann
Withering Syndrome (WS) is a lethal disease that affects abalone species in both wild and farmed populations. This infection, caused by the rickettsial-like intracellular organism (RLO) Candidatus Xenohaliotis californiensis, can severely impair the normal development of affected animals, and ultimately, their survival. The most common line of action against the WS has been the use of antibiotics, specifically oxytetracycline (OTC), administered via intramuscular injection and per os via medicated feed. In the present study, we have assessed the effectiveness of OTC baths as therapeutic treatment for the control of the WS agent in H...
February 28, 2018: Journal of Invertebrate Pathology
Vijaya Sarangthem, Eun A Cho, Aena Yi, Sang Kyoon Kim, Byung-Heon Lee, Rang-Woon Park
Expression of various molecules on the surface of cancer cells compared to normal cells creates a platform for the generation of various drug vehicles for targeted therapy. Multiple interactions between ligands and their receptors mediated by targeting peptide-modified polymer could enable simultaneous delivery of a drug selectively to target tumor cells, thus limiting side effects resulting from non-specific drug delivery. In this study, we synthesized a novel tumor targeting system by using two key elements: (1) Bld-1 peptide (SNRDARRC), a recently reported bladder tumor targeting peptide identified by using a phage-displayed peptide library, and (2) ELP, a thermally responsive polypeptide...
March 1, 2018: Scientific Reports
V Samsoninkova, N L Venkatareddy, W Wagermaier, A Dallmann, H G Börner
Peptide-polymer conjugates are applied as interface stabilizers that are precisely tuned to recognize the surfaces of inorganic constituents in composites. A set of peptide sequences is usually selected through phage-display and a strategy is presented for the identification of the most effective sequences through the evaluation of secondary interactions, including not only surface binding but also solubility and self-aggregation tendency.
March 1, 2018: Soft Matter
Marion David, Pascaline Lécorché, Maxime Masse, Aude Faucon, Karima Abouzid, Nicolas Gaudin, Karine Varini, Fanny Gassiot, Géraldine Ferracci, Guillaume Jacquot, Patrick Vlieghe, Michel Khrestchatisky
Insufficient membrane penetration of drugs, in particular biotherapeutics and/or low target specificity remain a major drawback in their efficacy. We propose here the rational characterization and optimization of peptides to be developed as vectors that target cells expressing specific receptors involved in endocytosis or transcytosis. Among receptors involved in receptor-mediated transport is the LDL receptor. Screening complex phage-displayed peptide libraries on the human LDLR (hLDLR) stably expressed in cell lines led to the characterization of a family of cyclic and linear peptides that specifically bind the hLDLR...
2018: PloS One
Sa Dong, Zongyi Bo, Cunzheng Zhang, Jianguo Feng, Xianjin Liu
Single-chain variable fragment (scFv) is a kind of antibody that possess only one chain of the complete antibody while maintaining the antigen-specific binding abilities and can be expressed in prokaryotic system. In this study, scFvs against Cry1 toxins were screened out from an immunized mouse phage displayed antibody library, which was successfully constructed with capacity of 6.25 × 107  CFU/mL. Using the mixed and alternative antigen coating strategy and after four rounds of affinity screening, seven positive phage-scFvs against Cry1 toxins were selected and characterized...
February 26, 2018: Applied Microbiology and Biotechnology
Wenjing Zhong, Guanghui Li, Xiaolu Yu, Min Zhu, Likun Gong, Yakun Wan
Bt Cry1B toxin, a residue in insect-resistant transgenic plants, has been identified to be harmful to human health. Therefore, it is urgent to detect the Cry1B toxin level in each kind of transgenic plant. Nbs, with prominently unique physiochemical properties, are becoming more and more promising tools in the detection of target antigens. In this study, an immune phage display library that was of high quality was successfully constructed for the screening of Cry1B-specific Nbs with excellent specificity, affinity, and thermostable...
February 24, 2018: MicrobiologyOpen
Samuel B Pollock, Amy Hu, Yun Mou, Alexander J Martinko, Olivier Julien, Michael Hornsby, Lynda Ploder, Jarrett J Adams, Huimin Geng, Markus Müschen, Sachdev S Sidhu, Jason Moffat, James A Wells
Human cells express thousands of different surface proteins that can be used for cell classification, or to distinguish healthy and disease conditions. A method capable of profiling a substantial fraction of the surface proteome simultaneously and inexpensively would enable more accurate and complete classification of cell states. We present a highly multiplexed and quantitative surface proteomic method using genetically barcoded antibodies called phage-antibody next-generation sequencing (PhaNGS). Using 144 preselected antibodies displayed on filamentous phage (Fab-phage) against 44 receptor targets, we assess changes in B cell surface proteins after the development of drug resistance in a patient with acute lymphoblastic leukemia (ALL) and in adaptation to oncogene expression in a Myc-inducible Burkitt lymphoma model...
February 23, 2018: Proceedings of the National Academy of Sciences of the United States of America
Steven Shave, Stefan Mann, Joanna Koszela, Alastair Kerr, Manfred Auer
The design of highly diverse phage display libraries is based on assumption that DNA bases are incorporated at similar rates within the randomized sequence. As library complexity increases and expected copy numbers of unique sequences decrease, the exploration of library space becomes sparser and the presence of truly random sequences becomes critical. We present the program PuLSE (Phage Library Sequence Evaluation) as a tool for assessing randomness and therefore diversity of phage display libraries. PuLSE runs on a collection of sequence reads in the fastq file format and generates tables profiling the library in terms of unique DNA sequence counts and positions, translated peptide sequences, and normalized 'expected' occurrences from base to residue codon frequencies...
2018: PloS One
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