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antibody phage display

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https://www.readbyqxmd.com/read/28820969/a-disulfide-stabilized-human-vl-single-domain-antibody-library-is-a-source-of-soluble-and-highly-thermostable-binders
#1
Kevin A Henry, Hiba Kandalaft, Michael J Lowden, Martin A Rossotti, Henk van Faassen, Greg Hussack, Yves Durocher, Dae Young Kim, Jamshid Tanha
We have previously shown that incorporation of a second intradomain disulfide linkage into camelid VHH and human VH/VL single-domain antibodies confers increased thermostability. Here, we explored the effects of introducing an additional disulfide linkage, formed between Cys48 and Cys64 (Kabat numbering), into a phage-displayed synthetic human VL library. In comparison to an identical library bearing only the highly conserved Cys23-Cys88 disulfide linkage, the disulfide-stabilized VL library tolerated a similar degree of randomization but retained a higher level of functional diversity after selection with protein L...
August 15, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28818634/mining-na%C3%A3-ve-rabbit-antibody-repertoires-by-phage-display-for-monoclonal-antibodies-of-therapeutic-utility
#2
Haiyong Peng, Thomas Nerreter, Jing Chang, Junpeng Qi, Xiuling Li, Pabalu Karunadharma, Gustavo Martinez, Mohammad Fallahi, Jo Soden, Jim Freeth, Roger R Beerli, Ulf Grawunder, Michael Hudecek, Christoph Rader
Owing to their high affinities and specificities, rabbit monoclonal antibodies (mAbs) have demonstrated value and potential primarily as basic research and diagnostic reagents, but in some cases also as therapeutics. To accelerate access to rabbit mAbs bypassing immunization, we generated a large naïve rabbit antibody repertoire represented by a phage display library encompassing >10 billion independent antibodies in chimeric rabbit/human Fab format and validated it by next-generation sequencing. Panels of rabbit mAbs selected from this library against two emerging cancer targets, ROR1 and ROR2, revealed high diversity, affinity, and specificity...
August 14, 2017: Journal of Molecular Biology
https://www.readbyqxmd.com/read/28796894/subtractive-phage-display-selection-for-screening-and-identification-of-peptide-sequences-with-potential-use-in-serodiagnosis-of-paracoccidioidomycosis-caused-by-paracoccidioides-brasiliensis
#3
Leticia da Silva Portes, Erika Seki Kioshima, Zoilo Pires de Camargo, Wagner Luiz Batista, Patricia Xander
Paracoccidioidomycosis (PCM) is a systemic granulomatous disease endemic in Latin America whose aetiologic agents are the thermodimorphic fungi Paracoccidioides brasiliensis and Paracoccidioides lutzii. Despite technological advances, some problems have been reported for the fungal antigens used for serological diagnosis, and inconsistencies among laboratories have been reported. The use of synthetic peptides in the serological diagnosis of infectious diseases has proved to be a valuable strategy because in some cases, the reactions are more specific and sensitive...
August 10, 2017: Letters in Applied Microbiology
https://www.readbyqxmd.com/read/28789584/monoclonal-antibodies-technologies-for-early-discovery-and-engineering
#4
Patrick J Kennedy, Carla Oliveira, Pedro L Granja, Bruno Sarmento
Antibodies are essential in modern life sciences biotechnology. Their architecture and diversity allow for high specificity and affinity to a wide array of biochemicals. Combining monoclonal antibody (mAb) technology with recombinant DNA and protein expression links antibody genotype with phenotype. Yet, the ability to select and screen for high affinity binders from recombinantly-displayed, combinatorial libraries unleashes the true power of mAbs and a flood of clinical applications. The identification of novel antibodies can be accomplished by a myriad of in vitro display technologies from the proven (e...
August 8, 2017: Critical Reviews in Biotechnology
https://www.readbyqxmd.com/read/28783899/development-of-a-phage-chemiluminescent-enzyme-immunoassay-with-high-sensitivity-for-the-determination-of-imidaclothiz-in-agricultural-and-environmental-samples
#5
Yuan Ding, Xiude Hua, Nana Sun, Jiachuan Yang, Jiaqi Deng, Haiyan Shi, Minghua Wang
In this study, we isolated six phage-displayed peptides by biopanning phage-displayed peptide libraries on an immobilized anti-imidaclothiz monoclonal antibody. After analyzing the relative sensitivity of the individual phage-displayed peptides, we subsequently developed and optimized both a phage enzyme immunoassay (P-ELISA) and a phage chemiluminescent enzyme immunoassay (P-CLEIA) to improve the sensitivity and linear range of imidaclothiz assays. The P-CLEIA (50% inhibition concentration (IC50) of 0.86ngmL(-1), linear range of 0...
July 31, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28772027/escherichia-coli-surface-display-for-the-selection-of-nanobodies
#6
REVIEW
Valencio Salema, Luis Ángel Fernández
Nanobodies (Nbs) are the smallest functional antibody fragments known in nature and have multiple applications in biomedicine or environmental monitoring. Nbs are derived from the variable segment of camelid heavy chain-only antibodies, known as VHH. For selection, libraries of VHH gene segments from naïve, immunized animals or of synthetic origin have been traditionally cloned in E. coli phage display or yeast display systems, and clones binding the target antigen recovered, usually from plastic surfaces with the immobilized antigen (phage display) or using fluorescence-activated cell sorting (FACS; yeast display)...
August 3, 2017: Microbial Biotechnology
https://www.readbyqxmd.com/read/28756139/a-universal-phage-display-system-for-the-seamless-construction-of-fab-libraries
#7
Renae S Nelson, Philippe Valadon
The construction of Fab phage libraries requires the cloning of domains from both the light and the heavy chain of antibodies. Despite the advent of powerful strategies such as splicing-by-overlap extension PCR, obtaining high quality libraries with excellent coverage remains challenging. Here, we explored the use of type IIS restriction enzymes for the seamless cloning of Fab libraries. We analyzed human, murine and rabbit germline antibody repertoires and identified combinations of restriction enzymes that exhibit very few or no recognition sites in the antibody sequences...
July 26, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28749765/generation-and-characterization-of-antinonstructural-protein-1-monoclonal-antibodies-and-development-of-diagnostics-for-dengue-virus-serotype-2
#8
Yin-Liang Tang, I-Ju Liu, Pi-Chun Li, Chien-Yu Chiu, Chun-Yu Lin, Chung-Hao Huang, Yen-Hsu Chen, Chi-Yu Fu, Day-Yu Chao, Chwan-Chuen King, Han-Chung Wu
Dengue virus (DENV) circulates in tropical and subtropical areas around the world, where it causes high morbidity and mortality. There is no effective treatment of infection, with supportive care being the only option. Furthermore, early detection and diagnosis are important to facilitate clinical decisions. In this study, seven monoclonal antibodies (mAbs) recognizing nonstructural protein 1 (NS1) of DENV were generated by hybridoma techniques. These antibodies can be divided into two groups: serotype-specific (DB6-1, DB12-3, and DB38-1) and nonspecific (consisting of antibodies DB16-1, DB20-6, DB29-1, and DB41-2)...
July 24, 2017: American Journal of Tropical Medicine and Hygiene
https://www.readbyqxmd.com/read/28739133/an-enzyme-linked-immunosorbent-assay-for-the-detection-of-diacetyl-2-3-butanedione
#9
Lucia Marri, Anita M Jansson, Caspar E Christensen, Ole Hindsgaul
Diacetyl (2,3-butanedione) is an important metabolic marker of several cancers, as well as an important off-flavour component produced during fermentation. As a small molecule in a complex mixture with many other analytes, existing methods for identification and quantitation of diacetyl invariably involves a chromatographic separation step followed by signal integration with an appropriate stoichiometric detector. Here we demonstrate that the chemical reaction of diacetyl with a 1,2-phenylenediamine derivative yields a chemical adduct, 1,4-quinoxaline which can be conjugated on BSA...
July 21, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28738290/cell-targeting-peptides-as-smart-ligands-for-targeting-of-therapeutic-or-diagnostic-agents-a-systematic-review
#10
REVIEW
Ali Mousavizadeh, Ali Jabbari, Mohammad Akrami, Hassan Bardania
Cell targeting peptides (CTP) are small peptides which have high affinity and specificity to a cell or tissue targets. They are typically identified by using phage display and chemical synthetic peptide library methods. CTPs have attracted considerable attention as a new class of ligands to delivery specifically therapeutic and diagnostic agents, because of the fact they have several advantages including easy synthesis, smaller physical sizes, lower immunogenicity and cytotoxicity and their simple and better conjugation to nano-carriers and therapeutic or diagnostic agents compared to conventional antibodies...
July 13, 2017: Colloids and Surfaces. B, Biointerfaces
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
#11
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
#12
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
#13
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
#14
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
#15
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...
August 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
#16
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
#17
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
#18
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
#19
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
#20
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
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