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https://www.readbyqxmd.com/read/28445507/various-mutations-compensate-for-a-deleterious-lacz%C3%AE-insert-in-the-replication-enhancer-of-m13-bacteriophage
#1
Emily M Zygiel, Karen A Noren, Marta A Adamkiewicz, Richard J Aprile, Heather K Bowditch, Christine L Carroll, Maria Abigail S Cerezo, Adelle M Dagher, Courtney R Hebert, Lauren E Hebert, Gloria M Mahame, Stephanie C Milne, Kelly M Silvestri, Sara E Sutherland, Alexandria M Sylvia, Caitlyn N Taveira, David J VanValkenburgh, Christopher J Noren, Marilena Fitzsimons Hall
M13 and other members of the Ff class of filamentous bacteriophages have been extensively employed in myriad applications. The Ph.D. series of phage-displayed peptide libraries were constructed from the M13-based vector M13KE. As a direct descendent of M13mp19, M13KE contains the lacZα insert in the intergenic region between genes IV and II, where it interrupts the replication enhancer of the (+) strand origin. Phage carrying this 816-nucleotide insert are viable, but propagate in E. coli at a reduced rate compared to wild-type M13 phage, presumably due to a replication defect caused by the insert...
2017: PloS One
https://www.readbyqxmd.com/read/28445413/identification-of-highly-specific-scfvs-against-total-adiponectin-for-diagnostic-purposes
#2
Peter Wilton, Michael Steidel, Gabriele Krczal, Iris Hermanns, Andreas Pfützner, Alisa Konnerth, Kajohn Boonrod
Adiponectin is one of the most abundant adipokines secreted from adipose tissue. It acts as an endogenous insulin sensitizer and plasma concentrations are inversely correlated with obesity and metabolic syndrome. A decrease in plasma adiponectin levels normally indicates increased hormonal activity of the visceral lipid tissue, which is associated with decreased insulin sensitivity. It may therefore be considered a valuable biomarker for elucidating the underlying deteriorations resulting in type 2 diabetes and macrovascular disease...
April 26, 2017: Biology
https://www.readbyqxmd.com/read/28444391/dual-display-phage-selection-driven-by-co-engagement-of-two-targets-by-two-different-antibody-fragments
#3
Séverine Fagète, Ledicia Botas-Perez, Irène Rossito-Borlat, Kenneth Adea, Franck Gueneau, Ulla Ravn, François Rousseau, Marie Kosco-Vilbois, Nicolas Fischer, Oliver Hartley
Antibody phage display technology has supported the emergence of numerous therapeutic antibodies. The development of bispecific antibodies, a promising new frontier in antibody therapy, could be facilitated by new phage display approaches that enable pairs of antibodies to be co-selected based on co-engagement of their respective targets. We describe such an approach, making use of two complementary leucine zipper domains that heterodimerize with high affinity. Phagemids encoding a first antibody fragment (scFv) fused to phage coat protein via the first leucine zipper are rescued in bacteria expressing a second scFv fused to the second leucine zipper as a soluble periplasmic protein, so that it is acquired by phage during assembly...
April 24, 2017: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/28443097/large-diversity-of-functional-nanobodies-from-a-camelid-immune-library-revealed-by-an-alternative-analysis-of-next-generation-sequencing-data
#4
Pieter Deschaght, Ana Paula Vintém, Marc Logghe, Miguel Conde, David Felix, Rob Mensink, Juliana Gonçalves, Jorn Audiens, Yanik Bruynooghe, Rita Figueiredo, Diana Ramos, Robbe Tanghe, Daniela Teixeira, Liesbeth Van de Ven, Catelijne Stortelers, Bruno Dombrecht
Next-generation sequencing (NGS) has been applied successfully to the field of therapeutic antibody discovery, often outperforming conventional screening campaigns which tend to identify only the more abundant selective antibody sequences. We used NGS to mine the functional nanobody repertoire from a phage-displayed camelid immune library directed to the recepteur d'origine nantais (RON) receptor kinase. Challenges to this application of NGS include accurate removal of read errors, correct identification of related sequences, and establishing meaningful inclusion criteria for sequences-of-interest...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28434129/phage-displayed-peptides-that-mimic-epitopes-of-hepatitis-e-virus-capsid
#5
Osmany Larralde, Juraj Petrik
Hepatitis E is an emerging zoonotic infection of increasing public health threat for the UK, especially for immunosuppressed individuals. A human recombinant vaccine has been licensed only in China and is not clear whether it protects against hepatitis E virus (HEV) genotype 3, the most prevalent in Europe. The aim of this study was to use phage display technology as a tool to identify peptides that mimic epitopes of HEV capsid (mimotopes). We identified putative linear and conformational mimotopes using sera from Scottish blood donors that have the immunological imprint of past HEV infection...
April 22, 2017: Medical Microbiology and Immunology
https://www.readbyqxmd.com/read/28428153/development-of-polyol-responsive-antibody-mimetics-for-single-step-protein-purification
#6
Richard J Suderman, Daren A Rice, Shane D Gibson, Eric J Strick, David M Chao
The purification of functional proteins is a critical pre-requisite for many experimental assays. Immunoaffinity chromatography, one of the fastest and most efficient purification procedures available, is often limited by elution conditions that disrupt structure and destroy enzymatic activity. To address this limitation, we developed polyol-responsive antibody mimetics, termed nanoCLAMPs, based on a 16 kDa carbohydrate binding module domain from Clostridium perfringens hyaluronidase. nanoCLAMPs bind targets with nanomolar affinity and high selectivity yet release their targets when exposed to a neutral polyol-containing buffer, a composition others have shown to preserve quaternary structure and enzymatic activity...
April 17, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28428053/discrimination-between-ulcerative-colitis-and-crohn-s-disease-using-phage-display-identified-peptides-and-virus-mimicking-synthetic-nanoparticles
#7
Sonia Facchin, Liboria Digiglio, Renata D'Incà, Elisabetta Casarin, Elisa Dassie, Monica Dettin, Annj Zamuner, Andrea Buda, Michele De Boni, Duilio Della Libera, Alessandra D'Urso, Giacomo Carlo Sturniolo, Margherita Morpurgo
Discrimination between Crohn Disease (CD) and Ulcerative Colitis (UC) is an unsolved clinical issue in up to 10-15% of inflammatory bowel disease (IBD) patients. We developed a method capable to discriminate between the two clinical conditions by combining random peptide phage display screenings and nanoparticle (NP) based assays. We identified two peptides able to recognize selectively biopsies of inflamed mucosa of either CD or UC patients. The peptides were integrated into synthetic virus-mimicking nanoassemblies using a poly-avidin NP platform...
April 17, 2017: Nanomedicine: Nanotechnology, Biology, and Medicine
https://www.readbyqxmd.com/read/28415614/connexin40-controls-endothelial-activation-by-dampening-nfkb-activation
#8
Jean-Francois Denis, Ke Ludwig Scheckenbach, Anna Pfenniger, Merlijn J Meens, Rob Krams, Lucile Miquerol, Steven Taffet, Marc Chanson, Mario Delmar, Brenda R Kwak
Connexins are proteins forming gap junction channels for intercellular communication. Connexin40 (Cx40) is highly expressed by endothelial cells (ECs) of healthy arteries but this expression is lost in ECs overlying atherosclerotic plaques. Low/oscillatory shear stress observed in bends and bifurcations of arteries is atherogenic partly through activation of the pro-inflammatory NFκB pathway in ECs. In this study, we investigated the relation between shear stress, Cx40 and NFκB. Shear stress-modifying casts were placed around carotid arteries of mice expressing eGFP under the Cx40 promoter (Cx40+/eGFP)...
March 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/28414460/analysis-of-novel-interactions-between-components-of-the-selenocysteine-biosynthesis-pathway-sephs1-sephs2-sepsecs-and-secp43
#9
Flore Oudouhou, Bastien Casu, Arnold Steve Dopgwa Puemi, Jurgen Sygusch, Christian Baron
In mammalian cells, the incorporation of the 21st amino acid, selenocysteine, into proteins is guided by the Sec machinery. The function of this protein complex requires several protein-protein and protein-RNA interactions, leading to the incorporation of selenocysteine at UGA codons. It is guided by stem-loop structures localized in the 3' untranslated regions of the selenoprotein-encoding genes. Here, we conducted a global analysis of interactions between the Sec biosynthesis and incorporation components using a bioluminescence resonance energy transfer assay in mammalian cells that showed that selenocysteine synthase (SEPSECS), SECp43, and selenophosphate synthetases SEPHS1 and SEPHS2 form oligomers in eukaryotic cells...
April 20, 2017: Biochemistry
https://www.readbyqxmd.com/read/28400756/antibody-engineering-for-pursuing-a-healthier-future
#10
REVIEW
Abdullah F U H Saeed, Rongzhi Wang, Sumei Ling, Shihua Wang
Since the development of antibody-production techniques, a number of immunoglobulins have been developed on a large scale using conventional methods. Hybridoma technology opened a new horizon in the production of antibodies against target antigens of infectious pathogens, malignant diseases including autoimmune disorders, and numerous potent toxins. However, these clinical humanized or chimeric murine antibodies have several limitations and complexities. Therefore, to overcome these difficulties, recent advances in genetic engineering techniques and phage display technique have allowed the production of highly specific recombinant antibodies...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28393683/identification-and-characterization-of-strychnine-binding-peptide-using-phage-display-screening
#11
Fang Zhang, Min Wang, Zheng Qiu, Xiao-Meng Wang, Chun-Lei Xu, Xia Zhang
Phage display has been proved in drug development as a high-throughput method for identifying specific cellular target of different drugs. However, phage display remains an intricate process for insoluble small chemicals because of the difficulty of presenting the molecules to the phages without occupying or destroying the very limited functional groups. In the present study, with Strychnine (Stry) selected as a model compound, we have developed an alternative in vitro bio-panning method against insoluble suspension...
April 4, 2017: Protein and Peptide Letters
https://www.readbyqxmd.com/read/28388565/prokaryotic-expression-of-mlaa-34-and-generation-of-a-novel-human-scfv-against-mlaa-34-by-phage-display-technology
#12
Yang Zhang, Pengyu Zhang, Aili He, Yun Yang, Jianli Wang, Hui Zhang, Wanggang Zhang
MLAA-34 is a newly identified monocytic leukemia-associated antigen that is overexpressed in acute monocytic leukemia specifically, thus providing a novel target for the therapy of acute monocytic leukemia. In this study, we first expressed MLAA-34 protein in Escherichia coli (E.coli) BL21 (DE3) cells and purified it by nickel ion affinity chromatography with high purity (>90%). Then, MLAA-34 was used as antigen for biopanning anti-MLAA-34 single chain antibody fragment (ScFv) from a fully human ScFv library, and a high affinity ScFv named MA1 was selected by phage-ELISA...
March 28, 2017: Oncotarget
https://www.readbyqxmd.com/read/28386109/the-underlying-process-of-early-ecological-and-genetic-differentiation-in-a-facultative-mutualistic-sinorhizobium-meliloti-population
#13
Nicolás Toro, Pablo J Villadas, María Dolores Molina-Sánchez, Pilar Navarro-Gómez, José M Vinardell, Lidia Cuesta-Berrio, Miguel A Rodríguez-Carvajal
The question of how genotypic and ecological units arise and spread in natural microbial populations remains controversial in the field of evolutionary biology. Here, we investigated the early stages of ecological and genetic differentiation in a highly clonal sympatric Sinorhizobium meliloti population. Whole-genome sequencing revealed that a large DNA region of the symbiotic plasmid pSymB was replaced in some isolates with a similar synteny block carrying densely clustered SNPs and displaying gene acquisition and loss...
April 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28383245/exploring-the-chemistry-of-genetic-information-storage-and-propagation-through-polymerase-engineering
#14
Gillian Houlihan, Sebastian Arangundy-Franklin, Philipp Holliger
Nucleic acids are a distinct form of sequence-defined biopolymer. What sets them apart from other biopolymers such as polypeptides or polysaccharides is their unique capacity to encode, store, and propagate genetic information (molecular heredity). In nature, just two closely related nucleic acids, DNA and RNA, function as repositories and carriers of genetic information. They therefore are the molecular embodiment of biological information. This naturally leads to questions regarding the degree of variation from this seemingly ideal "Goldilocks" chemistry that would still be compatible with the fundamental property of molecular heredity...
April 6, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28374253/display-of-hiv-1-envelope-protein-on-lambda-phage-scaffold-as-a-vaccine-platform
#15
Jonelle L Mattiacio, Matt Brewer, Stephen Dewhurst
The generation of a strong antibody response to target antigens is a major goal for vaccine development. Here we describe the display of the human immunodeficiency virus (HIV) envelope spike protein (Env) on a virus-like scaffold provided by the lambda phage capsid. Phage vectors, in general, have advantages over mammalian virus vectors due to their genetic tractability, inexpensive production, suitability for scale-up, as well as their physical stability, making them an attractive vaccine platform.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28373671/targeted-rescue-of-cancer-associated-idh1-mutant-activity-using-an-engineered-synthetic-antibody
#16
Shahir S Rizk, Somnath Mukherjee, Akiko Koide, Shohei Koide, Anthony A Kossiakoff
We have utilized a high-diversity phage display library to engineer antibody fragments (Fabs) that can modulate the activity of the enzyme isocitrate dehydrogenase 1 (IDH1). We show that a conformation-specific Fab can reactivate an IDH1 mutant associated with brain tumors. The results show that this strategy is a first step towards developing "activator drugs" for a large number of genetic disorders where mutations have disrupted protein function.
April 3, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28371143/taxonomic-variability-and-functional-stability-in-microbial-communities-infected-by-phages
#17
Stilianos Louca, Michael Doebeli
Microbial communities can display large variation in taxonomic composition, yet this variation can coincide with stable metabolic functional structure and performance. The mechanisms driving the taxonomic variation within functional groups remain largely unknown. Biotic interactions, such as predation by phages, have been suggested as potential cause of taxonomic turnover, but the conditions for this scenario have not been rigorously examined. Further, it is unknown how predation by phages affects community function, and how these effects are modulated by functional redundancy in the communities...
April 3, 2017: Environmental Microbiology
https://www.readbyqxmd.com/read/28369551/a-novel-excision-selection-method-for-isolation-of-antibodies-binding-antigens-expressed-specifically-by-rare-cells-in-tissue-sections
#18
Simon Lykkemark, Ole Aalund Mandrup, Mads Bjørnkjær Jensen, Jesper Just, Peter Kristensen
There is a growing appreciation of single cell technologies to provide increased biological insight and allow development of improved therapeutics. The central dogma explains why single cell technologies is further advanced in studies targeting nucleic acids compared to proteins, as nucleic acid amplification makes experimental detection possible. Here we describe a novel method for single round phage display selection of antibody fragments from genetic libraries targeting antigens expressed by rare cells in tissue sections...
March 27, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28369144/ankrd54-preferentially-selects-bruton-s-tyrosine-kinase-btk-from-a-human-src-homology-3-sh3-domain-library
#19
Manuela O Gustafsson, Dara K Mohammad, Erkko Ylösmäki, Hyunseok Choi, Subhash Shrestha, Qing Wang, Beston F Nore, Kalle Saksela, C I Edvard Smith
Bruton's Tyrosine Kinase (BTK) is a cytoplasmic protein tyrosine kinase with a fundamental role in B-lymphocyte development and activation. The nucleocytoplasmic shuttling of BTK is specifically modulated by the Ankyrin Repeat Domain 54 (ANKRD54) protein and the interaction is known to be exclusively SH3-dependent. To identify the spectrum of the ANKRD54 SH3-interactome, we applied phage-display screening of a library containing all the 296 human SH3 domains. The BTK-SH3 domain was the prime interactor. Quantitative western blotting analysis demonstrated the accuracy of the screening procedure...
2017: PloS One
https://www.readbyqxmd.com/read/28367456/synthetic-peptides-as-potential-antigens-for-cutaneous-leishmaniosis-diagnosis
#20
Juliana Seger Link, Silvana Maria Alban, Carlos Ricardo Soccol, Gilberto Vinicius Melo Pereira, Vanete Thomaz Soccol
This work's goal was to research new candidate antigens for cutaneous leishmaniosis (CL). In order to reach the goal, we used random peptide phage display libraries screened using antibodies from Leishmania braziliensis patients. After selection, three peptides (P1, P2, and P3) were synthesized using Fmoc chemistry. The peptides individually or a mixture of them (MIX) was subsequently emulsified in complete and incomplete Freund's adjuvant and injected subcutaneously in golden hamsters. Sera from the hamsters administered with P1 presented antibodies that recognized proteins between 76 and 150 kDa from L...
2017: Journal of Immunology Research
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