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https://www.readbyqxmd.com/read/27916124/alkaline-phosphatase-fused-repebody-as-a-new-format-of-immuno-reagent-for-an-immunoassay
#1
Hyo-Deok Seo, Joong-Jae Lee, Yu Jung Kim, Oliver Hantschel, Seung-Goo Lee, Hak-Sung Kim
Enzyme-linked immunoassays based on an antibody-antigen interaction are widely used in biological and medical sciences. However, the conjugation of an enzyme to antibodies needs an additional chemical process, usually resulting in randomly cross-linked molecules and a loss of the binding affinity and enzyme activity. Herein, we present the development of an alkaline phosphatase-fused repebody as a new format of immuno-reagent for immunoassays. A repebody specifically binding to human TNF-α (hTNF-α) was selected through a phage display, and its binding affinity was increased up to 49 nM using a modular engineering approach...
January 15, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/27915977/a-review-on-recent-patents-and-applications-of-inorganic-material-binding-peptides
#2
Veeranjaneyulu Thota, Carole C Perry
Over the past decade, significant progress has been made in the identification of novel material binding peptides having affinity to a wide range of target materials and their use in nanobiotechnological innovations. These material binding peptides (MBPs), also known as solid/ substance binding peptides (SBPs) can be isolated using combinatorial display technologies such as phage display (PD), surface display (cell, bacterial, yeast, mRNA) exhibit material specific selectivity and affinity towards a range of inorganic and organic nanomaterial surfaces including metals, metal oxides, minerals, semiconductors and biomolecules...
December 1, 2016: Recent Patents on Nanotechnology
https://www.readbyqxmd.com/read/27914072/identifying-bacterial-immune-evasion-proteins-using-phage-display
#3
Cindy Fevre, Lisette Scheepmaker, Pieter-Jan Haas
Methods aimed at identification of immune evasion proteins are mainly rely on in silico prediction of sequence, structural homology to known evasion proteins or use a proteomics driven approach. Although proven successful these methods are limited by a low efficiency and or lack of functional identification. Here we describe a high-throughput genomic strategy to functionally identify bacterial immune evasion proteins using phage display technology. Genomic bacterial DNA is randomly fragmented and ligated into a phage display vector that is used to create a phage display library expressing bacterial secreted and membrane bound proteins...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27906504/combination-of-two-epitope-identification-techniques-enables-the-rational-design-of-soy-allergen-gly-m-4-mutants
#4
Heide Havenith, Karolin Kern, Paul Rautenberger, Holger Spiegel, Michael Szardenings, Elke Ueberham, Jörg Lehmann, Matthias Buntru, Simon Vogel, Regina Treudler, Rainer Fischer, Stefan Schillberg
BACKGROUND: Detailed IgE-binding epitope analysis is a key requirement for the understanding and development of diagnostic and therapeutic agents to address food allergies. METHODS: We combined an IgE-specific linear peptide microarray with random phage peptide display for the high-resolution mapping of IgE-binding epitopes of the major soybean allergen Gly m 4, which is a homologue to the birch pollen allergen Bet v1. RESULTS: Three epitopes were identified and mapped to a resolution of four key amino acids, allowing the rational design and the production of three Gly m 4 mutants with the aim to abolish or reduce the binding of epitope-specific IgE...
December 1, 2016: Biotechnology Journal
https://www.readbyqxmd.com/read/27889295/novel-t7-like-expression-systems-used-for-halomonas
#5
Han Zhao, Haoqian M Zhang, Xiangbin Chen, Teng Li, Qiong Wu, Qi Ouyang, Guo-Qiang Chen
To engineer non-model organisms, suitable genetic parts must be available. However, biological parts are often host strain sensitive. It is therefore necessary to develop genetic parts that are functional regardless of host strains. Here we report several novel phage-derived expression systems used for transcriptional control in non-model bacteria. Novel T7-like RNA polymerase-promoter pairs were obtained by mining phage genomes, followed by in vivo characterization in non-model strains Halomonas spp TD01 and Pseudomonas entomophila...
November 24, 2016: Metabolic Engineering
https://www.readbyqxmd.com/read/27888111/neutralizing-human-recombinant-antibodies-against-herpes-simplex-virus-type-1-glycoproteins-b-from-a-phage-displayed-scfv-antibody-library
#6
Vahid Bagheri, Foroogh Nejatollahi, Seyed Alireza Esmaeili, Amir Abbas Momtazi, Mohamad Motamedifar, Amirhossein Sahebkar
The HSV-1 envelope glycoprotein B (gB) plays a critical role in virus entry into host cells. Neutralizing antibodies can therefore potentially prevent virus entry into target cells and cell-to-cell spread of infection. Our present study focused on the selection of neutralizing single-chain Fv (scFv) antibodies of a phage-displayed nonimmune human scFv antibody library against gB of HSV-1. To enrich specific scFvs, two phage antibodies were isolated against amino acid residues 31-43 derived from the N-terminal part of gB using panning technique...
November 22, 2016: Life Sciences
https://www.readbyqxmd.com/read/27887869/a-highly-diverse-and-functional-na%C3%A3-ve-ubiquitin-variant-library-for-generation-of-intracellular-affinity-reagents
#7
Isabel Leung, Nick Jarvik, Sachdev S Sidhu
We report the design, construction, and validation of a highly diverse phage-displayed naïve ubiquitin variant (Ubv) library. We first conducted a mutation tolerance scan of 27 residues and confirmed that 24 of these could be substituted by chemically diverse amino acids without compromising the display of Ubvs on phage. Subsequently, we constructed a library containing 6.8×10(10) unique members, in which these 24 positions were diversified with a degenerate codon that encodes for 6 aa that are prevalent in protein interaction sites...
November 22, 2016: Journal of Molecular Biology
https://www.readbyqxmd.com/read/27881076/clustering-of-disulfide-rich-peptides-provides-scaffolds-for-hit-discovery-by-phage-display-application-to-interleukin-23
#8
David T Barkan, Xiao-Li Cheng, Herodion Celino, Tran T Tran, Ashok Bhandari, Charles S Craik, Andrej Sali, Mark L Smythe
BACKGROUND: Disulfide-rich peptides (DRPs) are found throughout nature. They are suitable scaffolds for drug development due to their small cores, whose disulfide bonds impart extraordinary chemical and biological stability. A challenge in developing a DRP therapeutic is to engineer binding to a specific target. This challenge can be overcome by (i) sampling the large sequence space of a given scaffold through a phage display library and by (ii) panning multiple libraries encoding structurally distinct scaffolds...
November 23, 2016: BMC Bioinformatics
https://www.readbyqxmd.com/read/27879679/hydrostatin-tl1-an-anti-inflammatory-active-peptide-from-the-venom-gland-of-hydrophis-cyanocinctus-in-the-south-china-sea
#9
Ningyuan Wang, Yan Huang, An Li, Hailong Jiang, Jie Wang, Jianzhong Li, Lei Qiu, Ka Li, Yiming Lu
Tumor necrosis factor (TNF)-α is a pleiotropic cytokine with intense pro-inflammatory and immunomodulatory properties, and anti-TNF-α biologics are effective therapies for various inflammatory diseases such as inflammatory bowel disease (IBD) and sepsis. Snake venom, as a traditional Chinese medicine, has been used in the treatment of inflammatory diseases in China for centuries. In this research, we constructed a venom gland T7 phage display library of the sea snake Hydrophis cyanocinctus to screen bioactive compounds that antagonize TNF-α and identified a novel nine-amino-acid peptide, termed hydrostatin-TL1 (H-TL1)...
November 22, 2016: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/27874102/development-and-comparative-study-of-chemosynthesized-antigen-and-mimotope-based-immunoassays-for-class-specific-analysis-of-o-o-dimethyl-organophosphorus-pesticides
#10
Fengchun Zhao, Huimin Wang, Xiao Han, Zhengyou Yang
The multi-residue determination of organophosphorus pesticides (OPs) is an important task due to the wide application and high toxicity of OPs. However, there is no promising immunoassay to monitor the multi-residue of O,O-dimethyl OPs. In this study, a monoclonal antibody (mAb) against a generic hapten of O,O-dimethyl OPs (O,O-dimethyl O-(3-carboxyphenyl)phosphorothioate) was prepared. To develop an effective class-specific immunoassay, two strategies were performed to select the appropriate coating antigen or competing antigen...
November 22, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27873209/large-scale-interaction-profiling-of-protein-domains-through-proteomic-peptide-phage-display-using-custom-peptidomes
#11
Moon-Hyeong Seo, Satra Nim, Jouhyun Jeon, Philip M Kim
Protein-protein interactions are essential to cellular functions and signaling pathways. We recently combined bioinformatics and custom oligonucleotide arrays to construct custom-made peptide-phage libraries for screening peptide-protein interactions, an approach we call proteomic peptide-phage display (ProP-PD). In this chapter, we describe protocols for phage display for the identification of natural peptide binders for a given protein. We finally describe deep sequencing for the analysis of the proteomic peptide-phage display...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27873207/profiling-phosphopeptide-binding-domain-recognition-specificity-using-peptide-microarrays
#12
Michele Tinti, Simona Panni, Gianni Cesareni
Cellular organization and response to internal and external stimuli are mediated by an intricate web of protein interactions. Some of these interactions are regulated by covalent posttranslational modifications such as phosphorylation and acetylation. These modifications can change the chemical nature of the interaction interfaces and modulate the binding affinity of the interacting partners. In signal transduction, the most frequent modification is reversible phosphorylation of tyrosine, serine or threonine residues...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27867751/t7-phage-display-library-a-promising-strategy-to-detect-tuberculosis-specific-biomarkers
#13
Harvinder Talwar, Jaya Talreja, Lobelia Samavati
One-third of the world's population is infected with tuberculosis, only 10% will develop active disease and the remaining 90% is considered to have latent TB (LTB). While active TB is contagious and can be lethal, the LTB can evolve to active TB. The diagnosis of TB can be challenging, especially in the early stages, due to the variability in presentation and nonspecific signs and symptoms. Currently, we have limited tools available to diagnose active TB, predict treatment efficacy and cure of active tuberculosis, the reactivation of latent tuberculosis infection, and the induction of protective immune responses through vaccination...
June 2016: Mycobacterial Diseases: Tuberculosis & Leprosy
https://www.readbyqxmd.com/read/27864282/bxb1-integrase-serves-as-a-highly-efficient-dna-recombinase-in-rapid-metabolite-pathway-assembly
#14
Xianwei Wang, Biao Tang, Yu Ye, Yayi Mao, Xiaolai Lei, Guoping Zhao, Xiaoming Ding
Phage-encoded serine integrases are widely used in genetic engineering. They also have the potential to serve as efficient DNA assemblers, demonstrated by the method of site-specific recombination-based tandem assembly (SSRTA) that can combine biological parts into devices, pathways, and systems. Here, four serine integrases, ϕBT1, TG1, ϕRv1, and Bxb1, were investigated to ascertain their in vitro DNA assembly activities. Bxb1 integrase displayed the highest efficiency to obtain final products. Thus, we conclude that Bxb1 integrase is an excellent choice for DNA assembly in vitro Using this enzyme and its recognition sites, BioBrick standards were designed that are compatible with the SSRTA method for module addition...
November 17, 2016: Acta Biochimica et Biophysica Sinica
https://www.readbyqxmd.com/read/27863546/identification-of-three-novel-b-cell-epitopes-of-vmh-protein-from-vibrio-mimicus-by-screening-a-phage-display-peptide-library
#15
Ning Xiao, Ji Cao, Hao Zhou, Shu-Quan Ding, Ling-Yan Kong, Jin-Nian Li
Vibrio mimicus is the causative agent of ascites disease in fish. The heat-labile hemolytic toxin designated VMH is an immunoprotective antigen of V. mimicus. However, its epitopes have not been well characterized. Here, a commercially available phage displayed 12-mer peptide library was used to screen epitopes of VMH protein using polyclonal rabbit anti-rVMH protein antibodies, and then five positive phage clones were identified by sandwich and competitive ELISA. Sequences analysis showed that the motif of DPTLL displayed on phage clone 15 and the consensus motif of SLDDDST displayed on the clone 4/11 corresponded to the residues 134-138 and 238-244 of VMH protein, respectively, and the synthetic motif peptides could also be recognized by anti-rVMH-HD antibody in peptide-ELISA...
December 2016: Veterinary Immunology and Immunopathology
https://www.readbyqxmd.com/read/27863402/a-fully-human-anti-cd47-blocking-antibody-with-therapeutic-potential-for-cancer
#16
Dadi Zeng, Qiang Sun, Ang Chen, Jiangfeng Fan, Xiaopeng Yang, Lei Xu, Peng Du, Weiyi Qiu, Weicai Zhang, Shuang Wang, Zhiwei Sun
CD47/SIRPα interaction serves as an immune checkpoint for macrophage-mediated phagocytosis. Mouse anti-CD47 blocking antibodies had demonstrated potent efficacy in the treatment of both leukemic and solid tumors in preclinical experimentations, and therefore had moved forward rapidly into clinical trials. However, a fully human blocking antibody, which meets clinical purpose better, has not been reported for CD47 up to date. In this study, we reported the isolation of a fully human anti-CD47 blocking antibody, ZF1, from a phage display library...
November 15, 2016: Oncotarget
https://www.readbyqxmd.com/read/27862961/identification-of-a-glypican-3-binding-peptide-for-in-vivo-non-invasive-human-hepatocellular-carcinoma-detection
#17
Zainen Qin, Jingjing Wang, Ye Wang, Guohao Wang, Xiangyu Wang, Zhiyang Zhou, Gang Liu, Shi Gao, Lei Zhu
Early and accurate detection of hepatocellular carcinoma (HCC) is essential to improve the prognosis of patients and reduce the morbidity of surgical therapy. Glypican-3 (GPC3) is a protein abnormally expressed in HCC that has been identified as a serological and histochemical HCC marker. A novel peptide that specifically recognizes GPC3 will facilitate early detection of HCC and guide the treatment strategy. Herein, phage display screening technology is utilized to obtain a GPC3 binding peptide (GBP) using HCC cells expressing GPC3 in varying abundances...
November 15, 2016: Macromolecular Bioscience
https://www.readbyqxmd.com/read/27862752/improving-the-binding-affinity-of-in-vitro-evolved-cyclic-peptides-by-inserting-atoms-into-the-macrocycle-backbone
#18
Jonas Wilbs, Simon J Middendorp, Christian Heinis
Cyclic peptides binding to targets of interest can be generated efficiently with powerful in vitro display techniques, such as phage display or mRNA display. The cyclic peptide libraries screened with these methods are generated by altering in a combinatorial fashion the amino acid sequence of the peptides, the number of amino acids in the macrocycle rings, and the cyclization chemistry. A structural element that cannot easily be varied in the cyclic peptides is the backbone, which is built from amino acids, each of which contributes three atoms to the macrocyclic ring structure...
November 11, 2016: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/27861731/identification-of-mineral-binding-peptides-that-discriminate-between-chalcopyrite-and-enargite
#19
Susan Curtis, Franziska L Lederer, W Scott Dunbar, Ross T A MacGillivray
Innovative approaches to the separation of minerals and subsequent extraction of metals are imperative owing to the increasing mineralogical complexity of ore deposits that are difficult or even impossible to separate into slurries or solutions containing only the minerals or metals of interest. Low recovery of metal is typical for these complex deposits leading to significant losses to tailings. In addition, the minerals often contain impurities, some toxic, which are difficult and costly to control or manage during the processing of a concentrate or other mineral product...
November 14, 2016: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/27856318/sympathetic-nerve-repulsion-inhibited-by-designer-molecules-in-vitro-and-role-in-experimental-arthritis
#20
Julia Kunath, Nicolas Delaroque, Michael Szardenings, Ines Neundorf, Rainer H Straub
AIMS: In rheumatoid arthritis and collagen type II arthritis (CIA), sympathetic nerve fibers get lost in inflamed tissue. The process is probably induced by nerve repellent factors like semaphorin 3F (SEMA3F). Repulsion of sympathetic nerve fibers in inflamed tissue has proinflammatory effects due to the loss of anti-inflammatory neurotransmitters. We hypothesized that design molecules like antibodies and specific peptides that inhibit nerve fiber repulsion can ameliorate CIA. MATERIALS AND METHODS: Two blocking antibodies were used and four blocking peptides were generated using the phage display technique with the targets of SEMA3F and plexin-A2...
November 14, 2016: Life Sciences
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