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antibody phage display

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https://www.readbyqxmd.com/read/28642239/a-novel-single-domain-antibody-against-von-willebrand-factor-a1-domain-resolves-leukocyte-recruitment-and-vascular-leakage-during-inflammation
#1
Gabriel Aymé, Frédéric Adam, Paulette Legendre, Amine Bazaa, Valérie Proulle, Cécile V Denis, Olivier D Christophe, Peter J Lenting
OBJECTIVE: von Willebrand factor (VWF) is crucial to hemostasis, but also plays a role in inflammatory processes. Unfortunately, no proper monoclonal antibodies to study VWF function in mice are currently available. We therefore aimed to generate single-domain antibodies (sdAbs) recognizing murine VWF and blocking its function in vivo. APPROACH AND RESULTS: Llama-derived sdAbs recognizing both human and murine VWF were isolated via phage display technology. One of them (designated KB-VWF-006) recognized the VWF A1 domain with picomolar affinity...
June 22, 2017: Arteriosclerosis, Thrombosis, and Vascular Biology
https://www.readbyqxmd.com/read/28630449/effect-of-dna-sequence-of-fab-fragment-on-yield-characteristics-and-cell-growth-of-e-coli
#2
Antti Kulmala, Tuomas Huovinen, Urpo Lamminmäki
Codon usage is one of the factors influencing recombinant protein expression. We were interested in the codon usage of an antibody Fab fragment gene exhibiting extreme toxicity in the E. coli host. The toxic synthetic human Fab gene contained domains optimized by the "one amino acid-one codon" method. We redesigned five segments of the Fab gene with a "codon harmonization" method described by Angov et al. and studied the effects of these changes on cell viability, Fab yield and display on filamentous phage using different vectors and bacterial strains...
June 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28629727/antigenicity-of-phage-clones-and-their-synthetic-peptides-for-the-serodiagnosis-of-canine-and-human-visceral-leishmaniasis
#3
Lourena E Costa, Beatriz C S Salles, Thaís T O Santos, Fernanda F Ramos, Mariana P Lima, Mayara I S Lima, Áquila S B Portela, Miguel A Chávez-Fumagalli, Mariana C Duarte, Daniel Menezes-Souza, Ricardo A Machado-de-Ávila, Julia A G Silveira, Danielle F Magalhães-Soares, Luiz Ricardo Goulart, Eduardo A F Coelho
In the Americas, Brazil is responsible by 90% of the cases registered of visceral leishmaniasis (VL), and Leishmania infantum is the most common parasite species responsible by disease in Brazilian dogs and humans. A precise diagnosis may allow to a faster and more effective treatment against the disease, which increases the possibility of cure, as well as to induce less toxic effects, due to a lower time exposition for the chemotherapeutics. In a previous study, two L. infantum mimotopes, B10 and C01 clones, were recognized by antibodies in VL dogs sera by a phage display technology, and were well-successfully evaluated as vaccine candidates against visceral and tegumentary leishmaniasis...
June 16, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28627567/dissecting-binding-of-a-%C3%AE-barrel-membrane-protein-by-phage-display
#4
Luz M Meneghini, Sarvind Tripathi, Marcus A Woodworth, Sudipta Majumdar, Thomas L Poulos, Gregory A Weiss
Membrane proteins (MPs) constitute a third of all proteomes, and contribute to a myriad of cellular functions including intercellular communication, nutrient transport and energy generation. For example, TonB-dependent transporters (TBDTs) in the outer membrane of Gram-negative bacteria play an essential role transporting iron and other nutrients into the bacterial cell. The inherently hydrophobic surfaces of MPs complicates protein expression, purification, and characterization. Thus, dissecting the functional contributions of individual amino acids or structural features through mutagenesis can be a challenging ordeal...
June 19, 2017: Molecular BioSystems
https://www.readbyqxmd.com/read/28622860/immunogenicity-of-t7-bacteriophage-nanoparticles-displaying-g-h-loop-of-foot-and-mouth-disease-virus-fmdv
#5
Hai Xu, Xi Bao, Yu Lu, Yamei Liu, Bihua Deng, Yiwei Wang, Yue Xu, Jibo Hou
Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals that causes severe economic losses worldwide. The G-H loop of the FMDV VP1 structural protein is the major neutralizing antigenic site. However, a fully protective G-H loop peptide vaccine requires the addition of promiscuous Th sites from a source outside VP1. Thus, we demonstrated the potential of T7 bacteriophage based nanoparticles displaying a genetically fused G-H loop peptide (T7-GH) as a FMDV vaccine candidate. Recombinant T7-GH phage was constructed by inserting the G-H loop coding region into the T7 Select 415-1b vector...
June 2017: Veterinary Microbiology
https://www.readbyqxmd.com/read/28621534/construction-of-an-immunized-rabbit-phage-display-library-for-selecting-high-activity-of-against-bacillus-thuringiensis-cry1f-toxin-single-chain-antibodies
#6
Chongxin Xu, Cunzheng Zhang, Jianfeng Zhong, Hui Hu, Shimin Luo, Xiaoqin Liu, Xiao Zhang, Yuan Liu, Xianjin Liu
In the present study, a Cry1F-immunized rabbit phage display library (6.96×108) was constructed for selecting high activity of anti-Cry1F toxin single-chain antibody (scFv) by bio-panning. Total 16 positive monoclonal phage scFvs were obtained after 4 rounds panning which identified by enzyme-linked immunoassay (ELISA), PCR and DNA sequencing. The most positive phage scFv (named RF4) was expressed in E.coli.HB2151 and the purified soluble protein approximate 30 kDa with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)...
June 16, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28620550/generation-of-recombinant-antibodies-against-the-beta-1-6-branched-beta-1-3-d-glucan-schizophyllan-from-immunized-mice-via-phage-display
#7
Jörn Josewski, Sabine Buchmeier, André Frenzel, Philip Tinnefeld, Stefan Dübel, Udo Rau
beta-(1,6)-Branched beta-(1,3)-D-glucans like schizophyllan from the basidiomycete Schizophyllum commune excite various immunostimulatory effects and have been clinically tested as adjuvants. Some of the glucans are also applicable in food or petrol industry due to their viscosity and temperature stability in aqueous solution. Antibodies against these glucans could be used as tool for analysis of glucan preparations or for further research of its bioactivity. Therefore, an immune phage display library was constructed from mice immunized with schizophyllan...
2017: Biotechnology Research International
https://www.readbyqxmd.com/read/28619527/expression-of-full-length-her2-protein-in-sf9-insect-cells-and-its-presentation-on-the-surface-of-budded-virus-like-particles
#8
Lisa Nika, Jakob Wallner, Dieter Palmberger, Krisztina Koczka, Karola Vorauer-Uhl, Reingard Grabherr
Biomarkers of cancer are often glycosylated membrane receptor proteins present on the cellular surface. In order to develop new antibodies for cancer diagnostics or treatment, it is a main pre-requisite that these target proteins are available in a native conformation. However, membrane receptor proteins are notoriously difficult to produce due to their hydrophobic nature and complex architecture. Here, we used the baculovirus-insect cell expression system to produce budded virus-like particles (VLPs) as the scaffold for the presentation of complex membrane proteins...
June 12, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28617221/identification-of-cancer-specific-motifs-in-mimotope-profiles-of-serum-antibody-repertoire
#9
Ekaterina Gerasimov, Alex Zelikovsky, Ion Măndoiu, Yurij Ionov
BACKGROUND: For fighting cancer, earlier detection is crucial. Circulating auto-antibodies produced by the patient's own immune system after exposure to cancer proteins are promising bio-markers for the early detection of cancer. Since an antibody recognizes not the whole antigen but 4-7 critical amino acids within the antigenic determinant (epitope), the whole proteome can be represented by a random peptide phage display library. This opens the possibility to develop an early cancer detection test based on a set of peptide sequences identified by comparing cancer patients' and healthy donors' global peptide profiles of antibody specificities...
June 7, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28613102/a-high-throughput-platform-for-population-reformatting-and-mammalian-expression-of-phage-display-libraries-to-enable-functional-screening-as-full-length-igg
#10
Xiaodong Xiao, Julie A Douthwaite, Yan Chen, Ben Kemp, Sara Kidd, Jennifer Percival-Alwyn, Alison Smith, Kate Goode, Bonnie Swerdlow, David Lowe, Herren Wu, William F Dall'Acqua, Partha S Chowdhury
Phage display antibody libraries are a rich resource for discovery of potential therapeutic antibodies. Single-chain variable fragment (scFv) libraries are the most common format due to the efficient display of scFv by phage particles and the ease by which soluble scFv antibodies can be expressed for high-throughput screening. Typically, a cascade of screening and triaging activities are performed, beginning with the assessment of large numbers of E. coli-expressed scFv, and progressing through additional assays with individual reformatting of the most promising scFv to full-length IgG...
June 14, 2017: MAbs
https://www.readbyqxmd.com/read/28611730/exploiting-nanobodies-in-the-detection-and-quantification-of-human-growth-hormone-via-phage-sandwich-enzyme-linked-immunosorbent-assay
#11
Hossam Murad, Jana Mir Assaad, Rasha Al-Shemali, Abdul Qader Abbady
BACKGROUND: Monitoring blood levels of human growth hormone (hGH) in most children with short stature deficiencies is crucial for taking a decision of treatment with extended course of daily and expensive doses of recombinant hGH (rhGH or Somatropin(®)). Besides, misusing of rhGH by sportsmen is banned by the World Anti-Doping Agency and thus sensitive GH-detecting methods are highly welcome in this field. Nanobodies are the tiniest antigen-binding entity derived from camel heavy chain antibodies...
2017: Frontiers in Endocrinology
https://www.readbyqxmd.com/read/28597195/identification-of-epitopes-on-nonstructural-protein-7-of-porcine-reproductive-and-respiratory-syndrome-virus-recognized-by-monoclonal-antibodies-using-phage-display-technology
#12
Heng Wang, Rongchang Liu, Weidong Zhang, Lingshuang Sun, Zhangyong Ning, Fangxiao Ji, Jin Cui, Guihong Zhang
Nonstructural protein 7 (nsp7) of porcine reproductive and respiratory syndrome virus (PRRSV) is considered to be a suitable reagent for the development of serological diagnostic assays. It can be expressed as a soluble recombinant protein in Escherichia coli, and its antibody response may continue up to 202 days post-infection. Furthermore, the region encoded by nsp7 is highly homologous among various strains within the genotype, and the results of nsp7-based enzyme-linked immunosorbent assay (ELISA) showed high agreement with previous Idexx ELISA results...
June 8, 2017: Virus Genes
https://www.readbyqxmd.com/read/28596941/highly-specific-and-effective-targeting-of-egfrviii-positive-tumors-with-tandab-antibodies
#13
Kristina Ellwanger, Uwe Reusch, Ivica Fucek, Stefan Knackmuss, Michael Weichel, Thorsten Gantke, Vera Molkenthin, Eugene A Zhukovsky, Michael Tesar, Martin Treder
To harness the cytotoxic capacity of immune cells for the treatment of solid tumors, we developed tetravalent, bispecific tandem diabody (TandAb) antibodies that recognize EGFRvIII, the deletion variant III of the epidermal growth factor receptor (EGFR), and CD3 on T-cells, thereby directing immune cells to eliminate EGFRvIII-positive tumor cells. Using phage display, we identified scFv antibodies selectively binding to EGFRvIII. These highly EGFRvIII-specific, fully human scFv were substantially improved by affinity maturation, achieving KDs in the picomolar range, and were used to construct a set of bispecific EGFRvIII-targeting TandAbs with a broad range of binding and cytotoxic properties...
2017: Frontiers in Oncology
https://www.readbyqxmd.com/read/28592267/a-vaccine-with-a%C3%AE-oligomer-specific-mimotope-attenuates-cognitive-deficits-and-brain-pathologies-in-transgenic-mice-with-alzheimer-s-disease
#14
Shao-Wei Wang, Dong-Qun Liu, Ling-Xiao Zhang, Mei Ji, Yang-Xin Zhang, Quan-Xiu Dong, Shu-Ying Liu, Xi-Xiu Xie, Rui-Tian Liu
BACKGROUND: β-Amyloid peptide (Aβ) oligomers are initial factors used to induce Alzheimer's disease (AD) development, and Aβ monomers have normal physiological function. The antibodies or vaccines against Aβ monomers have serious problems, such as side effects and low curative effects. Therefore, it is essential to specifically target Aβ oligomers rather than monomers for the treatment of AD. METHODS: The mimotopes of Aβ oligomers were obtained by panning the phage-displayed random peptide libraries using oligomer-specific antibodies as targets and expressed on the surface of EBY100 Saccharomyces cerevisiae to generate yeast cell base vaccines...
June 7, 2017: Alzheimer's Research & Therapy
https://www.readbyqxmd.com/read/28591728/generation-and-evaluation-of-antibody-agents-for-molecular-imaging-of-cd44v6-expressing-cancers
#15
Anna-Karin Haylock, Johan Nilvebrant, Anja Mortensen, Irina Velikyan, Marika Nestor, Ronny Falk
AIM: The aim of this study was to generate and characterize scFv antibodies directed to human CD44v6, as well as to radiolabel and evaluate top candidates in vitro and in vivo for their potential use in CD44v6-targeted molecular imaging in cancer patients. MATERIALS AND METHODS: Phage display selections were used to isolate CD44v6-specific scFvs. A chain shuffling strategy was employed for affinity maturation based on a set of CD44v6-specific first-generation clones...
May 18, 2017: Oncotarget
https://www.readbyqxmd.com/read/28585908/tuning-the-specificity-of-a-two-in-one-fab-against-three-angiogenic-antigens-by-fully-utilizing-the-information-of-deep-mutational-scanning
#16
Patrick Koenig, Sarah Sanowar, Chingwei V Lee, Germaine Fuh
Monoclonal antibodies developed for therapeutic or diagnostic purposes need to demonstrate highly defined binding specificity profiles. Engineering of an antibody to enhance or reduce binding to related antigens is often needed to achieve the desired biological activity without safety concern. Here, we describe a deep sequencing-aided engineering strategy to fine-tune the specificity of an angiopoietin-2 (Ang2)/vascular endothelial growth factor (VEGF) dual action Fab, 5A12.1 for the treatment of age-related macular degeneration...
June 6, 2017: MAbs
https://www.readbyqxmd.com/read/28584111/phage-display-and-kinetic-selection-of-antibodies-that-specifically-inhibit-amyloid-self-replication
#17
Anna Munke, Jonas Persson, Tanja Weiffert, Erwin De Genst, Georg Meisl, Paolo Arosio, Anna Carnerup, Christopher M Dobson, Michele Vendruscolo, Tuomas P J Knowles, Sara Linse
The aggregation of the amyloid β peptide (Aβ) into amyloid fibrils is a defining characteristic of Alzheimer's disease. Because of the complexity of this aggregation process, effective therapeutic inhibitors will need to target the specific microscopic steps that lead to the production of neurotoxic species. We introduce a strategy for generating fibril-specific antibodies that selectively suppress fibril-dependent secondary nucleation of the 42-residue form of Aβ (Aβ42). We target this step because it has been shown to produce the majority of neurotoxic species during aggregation of Aβ42...
June 20, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28582852/human-anti-lipopolysaccharid-lps-antibodies-against-legionella-with-high-species-specificity
#18
Philipp Kuhn, Stefanie Thiem, Michael Steinert, Duncan Purvis, Veronika Lugmayr, Ulrich Treutlein, Lutz Plobner, Robert-Matthias Leiser, Michael Hust, Stefan Dübel
Legionella are Gram-negative bacteria that are ubiquitously present in natural and man-made water reservoirs. When humans inhale aerosolized water contaminated with Legionella, alveolar macrophages can be infected, which may lead to a life-threatening pneumonia called Legionnaires' disease. Due to the universal distribution of Legionella in water and their potential threat to human health, the Legionella concentration in water for human use must be strictly monitored, which is difficult since the standard detection still relies on lengthy cultivation and analysis of bacterial morphology...
May 8, 2017: Human Antibodies
https://www.readbyqxmd.com/read/28578824/improving-the-malaria-transmission-blocking-activity-of-a-plasmodium-falciparum-48-45-based-vaccine-antigen-by-spytag-spycatcher-mediated-virus-like-display
#19
Susheel K Singh, Susan Thrane, Christoph M Janitzek, Morten A Nielsen, Thor G Theander, Michael Theisen, Ali Salanti, Adam F Sander
Malaria is a devastating disease caused by Plasmodium parasites, resulting in almost 0.5 million deaths per year. The Pfs48/45 protein exposed on the P. falciparum sexual stages is one of the most advanced antigen candidates for a transmission-blocking (TB) vaccine in the clinical pipeline. However, it remains essential to identify an optimal vaccine formulation that can facilitate induction of a long-lasting TB anti-Pfs48/45 response. Here we report on the development and evaluation of two Pfs48/45-based virus-like particle (VLP) vaccines generated using the AP205 SpyTag/Catcher VLP system...
May 31, 2017: Vaccine
https://www.readbyqxmd.com/read/28572662/discovery-of-a-polystyrene-binding-peptide-isolated-from-phage-display-library-and-its-application-in-peptide-immobilization
#20
Xu Qiang, Keyong Sun, Lijun Xing, Yifeng Xu, Hong Wang, Zhengpin Zhou, Juan Zhang, Fang Zhang, Bilgen Caliskan, Min Wang, Zheng Qiu
Phage peptide display is a powerful technique for discovery of various target-specific ligands. However, target-unrelated peptides can often be obtained and cause ambiguous results. Peptide PB-TUP has been isolated repeatedly in our laboratory on different targets and we conducted a research on PB-TUP phage to investigate their binding properties and rate of propagation. ELISA and phage recovery assay demonstrated that PB-TUP phage had a significant superior affinity to polystyrene solid surface compared with control phage clones...
June 1, 2017: Scientific Reports
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