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antibody phage display

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https://www.readbyqxmd.com/read/28726978/analysis-of-the-substrate-specificity-of-factor-vii-activating-protease-fsap-and-design-of-specific-and-sensitive-peptide-substrates
#1
Emrah Kara, Dipankar Manna, Geir Åge Løset, Eric L Schneider, Charles S Craik, Sandip Kanse
Factor VII (FVII) activating protease (FSAP) is a circulating serine protease that is likely to be involved in a number of disease conditions such as stroke, atherosclerosis, liver fibrosis, thrombosis and cancer. To date, no systematic information is available about the substrate specificity of FSAP. Applying phage display and positional scanning substrate combinatorial library (PS-SCL) approaches we have characterised the specificity of FSAP towards small peptides. Results were evaluated in the context of known protein substrates as well as molecular modelling of the peptides in the active site of FSAP...
July 20, 2017: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/28725057/efficient-generation-of-single-domain-antibodies-with-high-affinities-and-enhanced-thermal-stabilities
#2
Naoya Shinozaki, Ryuji Hashimoto, Kiichi Fukui, Susumu Uchiyama
Single domain antibodies (sdAbs), made of natural single variable regions of camelid or cartilaginous fish antibodies, or unpaired variable regions of mouse or human IgGs, are some of the more promising biologic modalities. However, such conventional sdAbs have difficulties of either using unwieldy animals for immunization or having high affinity deficiencies. Herein, we offer a versatile method to generate rabbit variable domain of heavy chain (rVH) derived sdAbs with high affinities (K D values of single digit nM or less) and enhanced thermal stabilities (equal to or even higher than those of camelid derived sdAbs)...
July 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28719653/isolation-of-a-monoclonal-antibody-from-a-phage-display-library-binding-the-rhesus-macaque-mhc-class-i-allomorph-mamu-a1-001
#3
Nathan Holman, Jason T Weinfurter, Trevor R Harsla, Roger W Wiseman, Aaron J Belli, Anthony J Michaels, Keith A Reimann, Robert I DeMars, Matthew R Reynolds
Monoclonal antibodies that bind to human leukocyte antigen (HLA) are useful tools for HLA-typing, tracking donor-recipient chimerisms after bone marrow transplants, and characterizing specific major histocompatibility complexes (MHC) on cell surfaces. Unfortunately, equivalent reagents are not available for rhesus macaques, which are commonly used animal as models in organ transplant and infectious disease research. To address this deficiency, we isolated an antibody that recognizes the common Indian rhesus macaque MHC class I molecule, Mamu-A1*001...
2017: PloS One
https://www.readbyqxmd.com/read/28711695/identification-of-a-conformational-neutralizing-epitope-on-the-vp1-protein-of-type-a-foot-and-mouth-disease-virus
#4
Wenming Liu, Baolin Yang, Mingxia Wang, Haiwei Wang, Decheng Yang, Wenge Ma, Guohui Zhou, Li Yu
Foot-and-mouth disease (FMD) caused by foot-and-mouth disease virus (FMDV), is a highly contagious infectious disease that affects domestic and wild cloven-hoofed animals worldwide. In recent years, outbreaks of serotype A FMD have occurred in many countries. High-affinity neutralizing antibodies against a conserved epitope could provide protective immunity against diverse subtypes of FMDV serotype A and protect against future pandemics. In this study, we generated a serotype A FMDV-specific potent neutralizing monoclonal antibody (MAb), 6C9, which recognizes a conformation-dependent epitope...
July 8, 2017: Research in Veterinary Science
https://www.readbyqxmd.com/read/28690185/selection-of-similar-single-domain-antibodies-from-two-immune-vhh-libraries-obtained-from-two-alpacas-by-using-different-selection-methods
#5
Tengfei Li, Matthias Vandesquille, Sylvie Bay, Marc Dhenain, Benoît Delatour, Pierre Lafaye
The two most used methods to select camelid single-domain antibody-fragments (VHHs) are: displaying their repertoires on the surface of filamentous bacteriophages (phage display) or linking them to ribosomes (ribosome display). In this study, we compared specific VHHs isolated from two different immune libraries coming from two different alpacas by using these two selection methods. Three anti-GFAP (glial fibrillary acidic protein) VHHs were derived from an immune library obtained by ribosome display after immunization of one alpaca with purified GFAP, a protein expressed by astroglial cells...
July 6, 2017: Immunology Letters
https://www.readbyqxmd.com/read/28683141/isolation-of-serotype-specific-antibodies-against-dengue-virus-non-structural-protein-1-using-phage-display-and-application-in-a-multiplexed-serotyping-assay
#6
Kebaneilwe Lebani, Martina L Jones, Daniel Watterson, Andrea Ranzoni, Renee J Traves, Paul R Young, Stephen M Mahler
The multidimensional nature of dengue virus (DENV) infections, which can be caused by four distinct serotypes of the virus, complicates the sensitivity of assays designed for the diagnosis of infection. Different viral markers can be optimally detected at different stages of infection. Of particular clinical importance is the early identification of infection, which is pivotal for disease management and the development of blood screening assays. Non-structural protein 1 (NS1) is an early surrogate marker of infection and its detection in serum coincides with detectable viraemia...
2017: PloS One
https://www.readbyqxmd.com/read/28669053/efficient-targeting-of-cd13-on-cancer-cells-by-the-immunotoxin-scfv13-eta-and-the-bispecific-scfv-13xds16
#7
Elena Grieger, Gerrit Gresch, Judith Niesen, Mira Woitok, Stefan Barth, Rainer Fischer, Rolf Fendel, Christoph Stein
PURPOSE: Treatment of cancer using standard chemotherapy still offers a poor prognosis combined with severe side effects. Novel antibody-based therapies have been shown to overcome low efficiency and lack of selectivity by targeting cancer-associated antigens, such as aminopeptidase CD13. METHODS: We isolated a high-affinity CD13-specific single-chain fragment variable (scFv13) from a phage display library of V-genes from mice immunized with soluble antigen. An immunotoxin comprising the scFv13 and a truncated version of the exotoxin A of Pseudomonas aeruginosa (ETA', scFv13-ETA') and a bispecific scFv targeting CD13 and CD16 simultaneously (bsscFv[13xds16]) was generated and investigated for their therapeutic potential...
July 1, 2017: Journal of Cancer Research and Clinical Oncology
https://www.readbyqxmd.com/read/28667526/isolation-of-natural-anti-fc%C3%AE%C2%B5ri%C3%AE-autoantibodies-from-healthy-donors
#8
Monique Vogel, Michael P Horn
Natural antibodies are defined as antibodies detected in a healthy individual without active immunization. These antibodies are specific for exoantigens, as well as for autoantigens, mostly without any pathogenic role. Most of the studies conducted with natural (auto-) antibodies have been performed using affinity purified antibodies from individual sera or polyclonal Ig-preparations such as Intravenous Ig (IVIg). For in-depth analysis of such autoantibodies affinity-purified Ig-preparations from healthy individuals are of no use, as they are oligoclonal or polyclonal...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28661364/recombinant-antibodies-for-academia-a-practical-approach
#9
Pierre Cosson, Oliver Hartley
After several decades of optimization, phage display technology enables the routine isolation and production of recombinant monoclonal antibodies in vitro. As such it has the potential to provide the academic community with a vast, inexpensive and renewable supply of well-characterized reagents, reducing bottlenecks in basic science, helping increase reproducibility of experiments, and phasing out the use of animals for production and discovery of antibodies. Yet the overwhelming majority of fundamental research laboratories still use incompletely characterized antibodies developed in animals...
December 21, 2016: Chimia
https://www.readbyqxmd.com/read/28659884/fungal-glucosylceramide-specific-camelid-single-domain-antibodies-are-characterized-by-broad-spectrum-antifungal-activity
#10
Barbara De Coninck, Peter Verheesen, Christine M Vos, Inge Van Daele, Miguel F De Bolle, Joao V Vieira, Marnix Peferoen, Bruno P A Cammue, Karin Thevissen
Chemical crop protection is widely used to control plant diseases. However, the adverse effects of pesticide use on human health and environment, resistance development and the impact of regulatory requirements on the crop protection market urges the agrochemical industry to explore innovative and alternative approaches. In that context, we demonstrate here the potential of camelid single domain antibodies (VHHs) generated against fungal glucosylceramides (fGlcCer), important pathogenicity factors. To this end, llamas were immunized with purified fGlcCer and a mixture of mycelium and spores of the fungus Botrytis cinerea, one of the most important plant pathogenic fungi...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28659622/determination-of-aspergillus-pathogens-in-agricultural-products-by-a-specific-nanobody-polyclonal-antibody-sandwich-elisa
#11
Ting Wang, Peiwu Li, Qi Zhang, Wen Zhang, Zhaowei Zhang, Tong Wang, Ting He
Aspergillus and its poisonous mycotoxins are distributed worldwide throughout the environment and are of particular interest in agriculture and food safety. In order to develop a specific method for rapid detection of Aspergillus flavus to forecast diseases and control aflatoxins, a nanobody, PO8-VHH, highly reactive to A. flavus was isolated from an immunized alpaca nanobody library by phage display. The nanobody was verified to bind to the components of extracellular and intracellular antigen from both A...
June 28, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28658672/identification-of-a-novel-hiv-1-neutralizing-antibody-from-a-crf07_bc-infected-chinese-donor
#12
Youxiang Sun, Yuanyuan Qiao, Yuanmei Zhu, Huihui Chong, Yuxian He
The identification of human monoclonal antibodies (mAbs) able to neutralize a broad spectrum of primary HIV-1 isolates is highly important for understanding the immune response of HIV-1 infection and developing vaccines and therapeutics. In this study, we isolated a novel human mAb termed Y498 from a phage display antibody library constructed with the PBMC samples of a CRF07_BC-infected Chinese donor whose sera exhibited broadly neutralizing activity. Y498 cross-reacted with diverse Env antigens and neutralized 30% of 70 tested HIV-1 isolates...
June 21, 2017: Oncotarget
https://www.readbyqxmd.com/read/28654662/generation-of-a-rabbit-single-chain-fragment-variable-scfv-antibody-for-specific-detection-of-bradyrhizobium-sp-doa9-in-both-free-living-and-bacteroid-forms
#13
Nguyen Xuan Vu, Natcha Pruksametanan, Witsanu Srila, Watcharin Yuttavanichakul, Kamonluck Teamtisong, Neung Teaumroong, Nantakorn Boonkerd, Panlada Tittabutr, Montarop Yamabhai
A simple and reliable method for the detection of specific nitrogen-fixing bacteria in both free-living and bacteroid forms is essential for the development and application of biofertilizer. Traditionally, a polyclonal antibody generated from an immunized rabbit was used for detection. However, the disadvantages of using a polyclonal antibody include limited supply and cross-reactivity to related bacterial strains. This is the first report on the application of phage display technology for the generation of a rabbit recombinant monoclonal antibody for specific detection and monitoring of nitrogen-fixing bacteria in both free-living form and in plant nodules...
2017: PloS One
https://www.readbyqxmd.com/read/28651892/construction-of-a-recombinant-full-length-membrane-associated-igg-library
#14
Zehua Sun, Shiqiang Lu, Zheng Yang, Jingjing Li, Mei-Yun Zhang
HIV/AIDS has become a global pandemic. Development of an effective HIV-1 vaccine eliciting broadly neutralizing monoclonal antibodies (bnmAbs) remains a big challenge. Before an effective vaccine comes out, passive treatment for prevention and protection of HIV-1 infection may alleviate the burden caused by the pandemic. Numerous bnmAbs have been isolated against different epitopes in HIV-1 envelope glycoprotein via phage/yeast display, EBV-immortalization, single cell sorting and micro neutralization. Recombinant antibody library with extended diversity and enlarged size of units are applicable by phage/yeast display and mammalian cell display for monoclonal antibody isolation...
June 24, 2017: Virus Research
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
#15
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
#16
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
#17
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
#18
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
#19
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
#20
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
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