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antibody phage display

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https://www.readbyqxmd.com/read/28107434/anti-mrka-monoclonal-antibodies-reveal-distinct-structural-and-antigenic-features-of-mrka
#1
Qun Wang, Yan Chen, Romana Cvitkovic, Meghan E Pennini, Chew Shun Chang, Mark Pelletier, Jessica Bonnell, Adem C Koksal, Herren Wu, William F Dall'Acqua, C Kendall Stover, Xiaodong Xiao
Antibody therapy against antibiotics resistant Klebsiella pneumoniae infections represents a promising strategy, the success of which depends critically on the ability to identify appropriate antibody targets. Using a target-agnostic strategy, we recently discovered MrkA as a potential antibody target and vaccine antigen. Interestingly, the anti-MrkA monoclonal antibodies isolated through phage display and hybridoma platforms all recognize an overlapping epitope, which opens up important questions including whether monoclonal antibodies targeting different MrkA epitopes can be generated and if they possess different protective profiles...
2017: PloS One
https://www.readbyqxmd.com/read/28105549/automated-antibody-de-novo-sequencing-and-its-utility-in-biopharmaceutical-discovery
#2
K Ilker Sen, Wilfred H Tang, Shruti Nayak, Yong J Kil, Marshall Bern, Berk Ozoglu, Beatrix Ueberheide, Darryl Davis, Christopher Becker
Applications of antibody de novo sequencing in the biopharmaceutical industry range from the discovery of new antibody drug candidates to identifying reagents for research and determining the primary structure of innovator products for biosimilar development. When murine, phage display, or patient-derived monoclonal antibodies against a target of interest are available, but the cDNA or the original cell line is not, de novo protein sequencing is required to humanize and recombinantly express these antibodies, followed by in vitro and in vivo testing for functional validation...
January 19, 2017: Journal of the American Society for Mass Spectrometry
https://www.readbyqxmd.com/read/28103321/derivative-of-extremophilic-50s-ribosomal-protein-l35ae-as-an-alternative-protein-scaffold
#3
Anna V Lomonosova, Andrei B Ulitin, Alexei S Kazakov, Tajib A Mirzabekov, Eugene A Permyakov, Sergei E Permyakov
Small antibody mimetics, or alternative binding proteins (ABPs), extend and complement antibody functionality with numerous applications in research, diagnostics and therapeutics. Given the superiority of ABPs, the last two decades have witnessed development of dozens of alternative protein scaffolds (APSs) for the design of ABPs. Proteins from extremophiles with their high structural stability are especially favorable for APS design. Here, a 10X mutant of the 50S ribosomal protein L35Ae from hyperthermophilic archaea Pyrococcus horikoshii has been probed as an APS...
2017: PloS One
https://www.readbyqxmd.com/read/28101463/development-of-a-novel-human-single-chain-antibody-against-egfrviii-antigen-by-phage-display-technology
#4
Leila Rahbarnia, Safar Farajnia, Hossein Babaei, Jafar Majidi, Bahman Akbari, Shiva Ahdi Khosroshahi
Purpose: EGFRvIII as the most common mutant variant of the epidermal growth factor receptor is resulting from deletion of exons 2-7 in the coding sequence and junction of exons 1 and 8 through a novel glycine residue. EGFRvIII is highly expressed in glioblastoma, carcinoma of the breast, ovary, and lung but not in normal cells. The aim of the present study was identification of a novel single chain antibody against EGFRvIII as a promising target for cancer therapy. Methods: In this study, a synthetic peptide corresponding to EGFRvIII protein was used for screening a naive human scFv phage library...
December 2016: Advanced Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/28095447/identification-and-characterization-of-single-chain-antibodies-that-specifically-bind-gi-noroviruses
#5
Amy M Hurwitz, Wanzhi Huang, Baijun Kou, Mary K Estes, Robert L Atmar, Timothy Palzkill
Norovirus infections commonly lead to outbreaks of acute gastroenteritis and spread quickly, resulting in many health and economic challenges prior to diagnosis. Rapid and reliable diagnostic tests are therefore essential to identify infections and to guide the appropriate clinical responses at the point-of-care. Existing tools, including RT-PCR and enzyme immunoassays, pose several limitations based on the significant time, equipment and expertise required to elicit results. Immunochromatographic assays available for use at the point-of-care have poor sensitivity and specificity, especially for genogroup I noroviruses, thus requiring confirmation of results with more sensitive testing methods...
2017: PloS One
https://www.readbyqxmd.com/read/28092036/creation-of-phosphotyrosine-superbinders-by-directed-evolution-of-an-sh2-domain
#6
Haiming Huang, Tomonori Kaneko, Sachdev S Sidhu, Shawn S C Li
Commercial antibodies raised against phosphotyrosine have been widely used as reagents to detect or isolate tyrosine-phosphorylated proteins from cellular samples. However, these antibodies are costly and are not amenable to in-house production in an academic lab setting. In this chapter, we describe a method to generate super-high affinity SH2 domains, dubbed the phosphotyrosine superbinders, by evolving a natural SH2 domain using the phage display technology. The superbinders are stable and can be easily produced in Escherichia coli in large quantities...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28090795/isolation-and-characterization-of-anti-c-met-single-chain-fragment-variable-scfv-antibodies
#7
Elmira Safaie Qamsari, Zahra Sharifzadeh, Salman Bagheri, Farhad Riazi-Rad, Vahid Younesi, Mohsen Abolhassani, Sepideh Safaei Ghaderi, Behzad Baradaran, Mohammad Hossein Somi, Mehdi Yousefi
The receptor tyrosine kinase (RTK) Met is the cell surface receptor for hepatocyte growth factor (HGF) involved in invasive growth programs during embryogenesis and tumorgenesis. There is compelling evidence suggesting important roles for c-Met in colorectal cancer proliferation, migration, invasion, angiogenesis, and survival. Hence, a molecular inhibitor of an extracellular domain of c-Met receptor that blocks c-Met-cell surface interactions could be of great thera-peutic importance. In an attempt to develop molecular inhibitors of c-Met, single chain variable fragment (scFv) phage display libraries Tomlinson I + J against a specific synthetic oligopeptide from the extracellular domain of c-Met receptor were screened; selected scFv were then characterized using various immune techniques...
January 16, 2017: Journal of Immunotoxicology
https://www.readbyqxmd.com/read/28075174/analysis-of-sivmac-envelope-specific-antibodies-selected-via-phage-display
#8
Sergio Ita, Mayara Rovariz Agostinho, Katherine Sullivan, Seung Yub Han, Rana Akleh, Welkin Johnson, Ismael Ben Farouck Fofana
We have constructed a single chain Fv (scFv) phage display library from an SIV-infected rhesus macaque that developed unusually high-titer neutralizing antibody responses against tier-3, neutralization-resistant SIVmac239. The library was screened using trimeric (gp140) and monomeric (gp120) forms of the SIVmac239 envelope (Env) glycoprotein. We also cloned variable-heavy and variable-light (VH-VL) antibody fragments from 7 previously described rhesus macaque B-cell lines (BLCL) that produce SIV gp120-specific monoclonal antibodies (mAbs)...
January 11, 2017: AIDS Research and Human Retroviruses
https://www.readbyqxmd.com/read/28075071/multi-target-selection-of-catalytic-antibodies-wih-%C3%AE-lactamase-activity-using-phage-display
#9
Melody A Shahsavarian, Nancy Chaaya, Narciso Costa, Didier Boquet, Alexandre Atkinson, Bernard Offmann, Srini V Kaveri, Sébastien Lacroix-Desmazes, Alain Friboulet, Bérangère Avalle, Séverine Padiolleau-Lefèvre
β-lactamase enzymes responsible for bacterial resistance to antibiotics are among the most important health threats to the human population today. Understanding the increasingly vast structural motifs responsible for the catalytic mechanism of β-lactamases will help improve the future design of new generation antibiotics and mechanism-based inhibitors of these enzymes. Here we report the construction of a large murine scFv phage display library of size 2.7×10(9) with extended diversity by combining different mouse models...
January 11, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28072528/triosephosphate-isomerase-and-filamin-c-share-common-epitopes-as-novel-allergens-of-procambarus-clarkii
#10
Yang Yang, Yong-Xia Zhang, Meng Liu, Soheila J Maleki, Ming-Li Zhang, Qingmei Liu, Min-Jie Cao, Wen-Jin Su, Guang-Ming Liu
Triosephosphate isomerase (TIM) is a key enzyme in glycolysis and has been identified as an allergen in saltwater products. In this study, TIM with the molecular mass of 28 kDa was purified from the freshwater crayfish (Procambarus clarkii) muscle. A 90-kDa protein showed IgG/IgE cross-reactivity with TIM was purified and identified as filamin C (FLN c), which is an actin-binding proteins. TIM showed similar thermal and pH stability while better digestion resistant compared with FLN c. Result of surface plasmon resonance (SPR) experiment demonstrated the infinity of anti-TIM polyclonal antibody (pAb) to both TIM and FLN c...
January 10, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28064157/fviii-specific-human-chimeric-antigen-receptor-car-t-regulatory-cells-suppress-t-and-b-cell-responses-to-fviii
#11
Jeongheon Yoon, Anja Schmidt, Ai-Hong Zhang, Christoph Königs, Yong Chan Kim, David W Scott
Replacement therapy with factor VIII (FVIII) is used in hemophilia A patients for treatment of bleeding episodes or for prophylaxis. A common and serious problem with this therapy is the patient's immune response to FVIII, due to a lack of tolerance, leading to the formation of inhibitory antibodies. Development of tolerogenic therapies, other than standard ITI, is an unmet goal. We previously generated engineered antigen- specific regulatory T cells (Tregs), created by transduction of a recombinant T cell receptor (TCR) isolated from a hemophilia A subject's T cell clone...
November 15, 2016: Blood
https://www.readbyqxmd.com/read/28062210/hyperthermostable-binding-molecules-on-phage-assay-components-for-point-of-care-diagnostics-for-active-tuberculosis-infection
#12
Ning Zhao, John Spencer, Margaret A Schmitt, John D Fisk
Tuberculosis is the leading cause of death from infectious disease worldwide. The low sensitivity, extended processing time, and high expense of current diagnostics are major challenges to the detection and treatment of tuberculosis. Mycobacterium tuberculosis ornithine transcarbamylase (Mtb OTC, Rv1656) has been identified in the urine of patients with active TB infection and is a promising target for point-of-care diagnostics. Specific binding proteins with low nanomolar affinities for Mtb OTC were selected from a phage display library built upon a hyperthermostable Sso7d scaffold...
January 3, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28056857/upregulation-of-mrps18a-in-breast-cancer-identified-by-selecting-phage-antibody-libraries-on-breast-tissue-sections
#13
Karen Marie Juul Sørensen, Theresa Meldgaard, Connie Jenning Melchjorsen, Agla J Fridriksdottir, Henrik Pedersen, Ole William Petersen, Peter Kristensen
BACKGROUND: One of the hallmarks of cancer is an altered energy metabolism, and here, mitochondria play a central role. Previous studies have indicated that some mitochondrial ribosomal proteins change their expression patterns upon transformation. METHOD: In this study, we have used the selection of recombinant antibody libraries displayed on the surface of filamentous bacteriophage as a proteomics discovery tool for the identification of breast cancer biomarkers...
January 5, 2017: BMC Cancer
https://www.readbyqxmd.com/read/28039694/removal-of-b-cell-epitopes-for-decreasing-immunogenicity-in-recombinant-immunotoxin-against-b-cell-malignancies
#14
Xiabo Hu, Min Zhang, Caiping Zhang, Shiyin Long, Wuzhou Wang, Weidong Yin, Zhaohui Cao
Recombinant immunotoxin HA22, composed of an anti- CD22 Fv fragment fused to PE38, a truncated portion of Pseudomonas Exotoxin A (PE), has been developed for targeted treatment of various B-cell malignancies. As a foreign, internalized macromolecule, PE38 often induces lysosomal degradation and neutralizing antibodies to limit the efficacy of treating B-cell malignancies. The region of PE38 containing lysosomal protease cleavage sites deleted, leaving only furin processing site. The resulting immunotoxin HA22-LR (lysosome resistant) retains excellent biologic activity and removes immunogenic epitopes as an additional benefit...
November 2016: Journal of B.U.ON.: Official Journal of the Balkan Union of Oncology
https://www.readbyqxmd.com/read/28017694/pitfalls-to-avoid-when-using-phage-display-for-snake-toxins
#15
Andreas Hougaard Laustsen, Line Præst Lauridsen, Bruno Lomonte, Mikael Rørdam Andersen, Brian Lohse
Antivenoms against bites and stings from snakes, spiders, and scorpions are associated with immunological side effects and high cost of production, since these therapies are still derived from the serum of hyper-immunized production animals. Biotechnological innovations within envenoming therapies are thus warranted, and phage display technology may be a promising avenue for bringing antivenoms into the modern era of biologics. Although phage display technology represents a robust and high-throughput approach for the discovery of antibody-based antitoxins, several pitfalls may present themselves when animal toxins are used as targets for phage display selection...
December 23, 2016: Toxicon: Official Journal of the International Society on Toxinology
https://www.readbyqxmd.com/read/28017498/red-cell-immunohematology-research-conducted-in-china
#16
REVIEW
Ziyan Zhu, Luyi Ye, Qin Li, Hongwei Gao, Yinxia Tan, Wei Cai
ABO subtypes and RhD variants are the most studied blood groups in China. Some of the polymorphisms in these two blood groups have direct clinical relevance. Molecular diagnosis of blood group polymorphisms is underway in China. In addition, research groups have developed methods such as screening for blood group mimetic peptides using phage display technology. New reagents, akin to antibodies directed against RhD and ABO, are being investigated using aptamer-based techniques. Progress is also being made in the development of synthetic exoglycosidases for conversion of group A and/or B antigens to group O...
November 24, 2016: Transfusion Medicine Reviews
https://www.readbyqxmd.com/read/28008969/determination-of-equilibrium-dissociation-constants-for-recombinant-antibodies-by-high-throughput-affinity-electrophoresis
#17
Yuchen Pan, Eric K Sackmann, Karolina Wypisniak, Michael Hornsby, Sammy S Datwani, Amy E Herr
High-quality immunoreagents enhance the performance and reproducibility of immunoassays and, in turn, the quality of both biological and clinical measurements. High quality recombinant immunoreagents are generated using antibody-phage display. One metric of antibody quality - the binding affinity - is quantified through the dissociation constant (KD) of each recombinant antibody and the target antigen. To characterize the KD of recombinant antibodies and target antigen, we introduce affinity electrophoretic mobility shift assays (EMSAs) in a high-throughput format suitable for small volume samples...
December 23, 2016: Scientific Reports
https://www.readbyqxmd.com/read/28004095/epitope-mapping-of-mono-and-polyclonal-antibodies-by-screening-phage-displayed-random-peptide-libraries
#18
Peter Molek, Tomaž Bratkovič
Detailed knowledge of antigenic determinants is crucial when characterizing therapeutic and diagnostic antibodies, assessing vaccine effectiveness and developing epitope-based vaccines. Most epitope mapping approaches are labor intensive and costly. In this study, we evaluated panning of phage-displayed random peptide libraries against antibodies as a tool for cognate epitope identification. We used six antibodies directed to three model protein antigens as targets to show that the approach is applicable to both mono- and polyclonal antibodies...
December 2016: Acta Chimica Slovenica
https://www.readbyqxmd.com/read/27989733/immunomagnetic-separation-of-human-myeloperoxidase-using-an-antibody-mimicking-peptide-identified-by-phage-display
#19
Soi Yun, Hyunmin Ryu, E K Lee
Phage display biopanning is a powerful in vitro selection process for screening and identifying peptides that bind to a target protein of interest. With the aim of replacing antibodies in immuno-diagnostic applications, we identified peptides whose binding characteristics mimicked those of anti-human myeloperoxidase (hMPO), a biomarker for acute cardiac diseases. Based on ELISA results from four phage clones, we selected and chemically synthesized a 12-mer peptide (SYIEPPERHRHR). Quartz crystal microbalance and surface plasmon resonance analyses revealed that the molar binding equilibrium ratio of the synthesized peptide was 0...
December 15, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27984065/isolation-and-characterization-of-a-novel-human-scfv-inhibiting-egfr-viii-expressing-cancers
#20
Leila Rahbarnia, Safar Farajnia, Hossein Babaei, Jafar Majidi, Hassan Dariushnejad, Mohammad Kazem Hosseini
EGFRvIII, a mutant form of epidermal growth factor receptor is highly expressed in glioblastoma, carcinoma of the breast, ovary, and lung but not in normal cells. This tumor specific antigen has emerged as a promising candidate for antibody based therapy of several cancers. The aim of the present study was isolation and characterization of a human single chain antibody against EGFRvIII as a promising target for cancer therapy. For this, a synthetic peptide corresponding to EGFRvIII protein was used for screening the naive human scFv phage library...
October 29, 2016: Immunology Letters
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