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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/28813669/the-phage-nucleus-and-tubulin-spindle-are-conserved-among-large-pseudomonas-phages
#3
Vorrapon Chaikeeratisak, Katrina Nguyen, MacKennon E Egan, Marcella L Erb, Anastasia Vavilina, Joe Pogliano
We recently demonstrated that the large Pseudomonas chlororaphis bacteriophage 201φ2-1 assembles a nucleus-like structure that encloses phage DNA and segregates proteins according to function, with DNA processing proteins inside and metabolic enzymes and ribosomes outside the nucleus. Here, we investigate the replication pathway of the Pseudomonas aeruginosa bacteriophages φKZ and φPA3. Bacteriophages φKZ and φPA3 encode a proteinaceous shell that assembles a nucleus-like structure that compartmentalizes proteins and DNA during viral infection...
August 15, 2017: Cell Reports
https://www.readbyqxmd.com/read/28811841/bacteriophages-in-the-gastrointestinal-tract-and-their-implications
#4
Marzanna Łusiak-Szelachowska, Beata Weber-Dąbrowska, Ewa Jończyk-Matysiak, Renata Wojciechowska, Andrzej Górski
The gut microbiota plays an essential role in health and disease of humans. Bacteriophages are the most abundant members of the gut virobiota and display great diversity. Phages can translocate through the mucosa to lymph and internal organs and play a role as regulators of the bacterial population in the gut. Increasing abundance of phages in the gut mucosa may reduce colonization by bacteria. Moreover, phages may have an immunomodulatory role in the immune response in the human gut. The role of phages in inflammatory bowel disease (IBD) remains unknown...
2017: Gut Pathogens
https://www.readbyqxmd.com/read/28811342/bacteriophages-of-gordonia-spp-display-a-spectrum-of-diversity-and-genetic-relationships
#5
Welkin H Pope, Travis N Mavrich, Rebecca A Garlena, Carlos A Guerrero-Bustamante, Deborah Jacobs-Sera, Matthew T Montgomery, Daniel A Russell, Marcie H Warner, Graham F Hatfull
The global bacteriophage population is large, dynamic, old, and highly diverse genetically. Many phages are tailed and contain double-stranded DNA, but these remain poorly characterized genomically. A collection of over 1,000 phages infecting Mycobacterium smegmatis reveals the diversity of phages of a common bacterial host, but their relationships to phages of phylogenetically proximal hosts are not known. Comparative sequence analysis of 79 phages isolated on Gordonia shows these also to be diverse and that the phages can be grouped into 14 clusters of related genomes, with an additional 14 phages that are "singletons" with no closely related genomes...
August 15, 2017: MBio
https://www.readbyqxmd.com/read/28808028/immune-evasion-by-a-staphylococcal-inhibitor-of-myeloperoxidase
#6
Nienke W M de Jong, Kasra X Ramyar, Fermin E Guerra, Reindert Nijland, Cindy Fevre, Jovanka M Voyich, Alex J McCarthy, Brandon L Garcia, Kok P M van Kessel, Jos A G van Strijp, Brian V Geisbrecht, Pieter-Jan A Haas
Staphylococcus aureus is highly adapted to its host and has evolved many strategies to resist opsonization and phagocytosis. Even after uptake by neutrophils, S. aureus shows resistance to killing, which suggests the presence of phagosomal immune evasion molecules. With the aid of secretome phage display, we identified a highly conserved protein that specifically binds and inhibits human myeloperoxidase (MPO), a major player in the oxidative defense of neutrophils. We have named this protein "staphylococcal peroxidase inhibitor" (SPIN)...
August 14, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28807301/a-candidate-recombinant-antigen-for-diagnosis-of-schistosomiasis-japonica-in-domestic-animals
#7
Jintao Feng, Rui Xu, Xin Zhang, Yu Han, Chuanchuan He, Chao Lu, Yang Hong, Ke Lu, Hao Li, Yamei Jin, Jiaojiao Lin, Jinming Liu
Domestic animals infected with Schistosoma japonicum are a major source of infection and play an important role in transmission to humans. A key strategy for the elimination of schistosomiasis is to control the sources of infection. In the present study, we identified a candidate diagnostic antigen-encoding gene, SjMRP1, the putative multidrug resistance protein 1 gene, by screening a cDNA phage display library from 44-day-old S. japonicum worms using IgGs from goat, cattle, and buffalo infected with S. japonicum...
August 30, 2017: Veterinary Parasitology
https://www.readbyqxmd.com/read/28797098/comparative-genomics-of-two-super-shedder-isolates-of-escherichia-coli-o157-h7
#8
Robab Katani, Rebecca Cote, Indira T Kudva, Chitrita DebRoy, Terrance M Arthur, Vivek Kapur
Shiga toxin-producing Escherichia coli O157:H7 (O157) are zoonotic foodborne pathogens and of major public health concern that cause considerable intestinal and extra-intestinal illnesses in humans. O157 colonize the recto-anal junction (RAJ) of asymptomatic cattle who shed the bacterium into the environment through fecal matter. A small subset of cattle, termed super-shedders (SS), excrete O157 at a rate (≥ 104 CFU/g of feces) that is several orders of magnitude greater than other colonized cattle and play a major role in the prevalence and transmission of O157...
2017: PloS One
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
#9
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
#10
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
#11
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/28780344/selection-of-affinity-peptides-for-interference-free-detection-of-cholera-toxin
#12
Jong Min Lim, Nam Su Heo, Seo Yeong Oh, Myung Yi Ryu, Jeong Hyun Seo, Tae Jung Park, Yun Suk Huh, Jong Pil Park
Cholera toxin is a major virulent agent of Vibrio cholerae, and it can rapidly lead to severe dehydration, shock, causing death within hours without appropriate clinical treatments. In this study, we present a method wherein unique and short peptides that bind to cholera toxin subunit B (CTX-B) were selected through M13 phage display. Biopanning over recombinant CTX-B led to rapid screening of a unique peptide with an amino acid sequence of VQCRLGPPWCAK, and the phage-displayed peptides analyzed using ELISA, were found to show specific affinities towards CTX-B...
July 31, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28773822/identification-of-specific-hydroxyapatite-001-binding-heptapeptide-by-phage-display-and-its-nucleation-effect
#13
Jing Mao, Xin Shi, Ya-Bo Wu, Shi-Qiang Gong
With recent developments of molecular biomimetics that combine genetic engineering and nanotechnology, peptides can be genetically engineered to bind specifically to inorganic components and execute the task of collagen matrix proteins. In this study, using biogenous tooth enamel as binding substrate, we identified a new heptapeptide (enamel high-affinity binding peptide, EHBP) from linear 7-mer peptide phage display library. Through the output/input affinity test, it was found that EHBP has the highest affinity to enamel with an output/input ratio of 14...
August 17, 2016: Materials
https://www.readbyqxmd.com/read/28772478/biogenic-and-synthetic-peptides-with-oppositely-charged-amino-acids-as-binding-sites-for-mineralization
#14
Marie-Louise Lemloh, Klara Altintoprak, Christina Wege, Ingrid M Weiss, Dirk Rothenstein
Proteins regulate diverse biological processes by the specific interaction with, e.g., nucleic acids, proteins and inorganic molecules. The generation of inorganic hybrid materials, such as shell formation in mollusks, is a protein-controlled mineralization process. Moreover, inorganic-binding peptides are attractive for the bioinspired mineralization of non-natural inorganic functional materials for technical applications. However, it is still challenging to identify mineral-binding peptide motifs from biological systems as well as for technical systems...
January 28, 2017: Materials
https://www.readbyqxmd.com/read/28772027/escherichia-coli-surface-display-for-the-selection-of-nanobodies
#15
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/28758361/igy-binding-peptide-screened-from-a-random-peptide-library-as-a-ligand-for-igy-purification
#16
Kamrul Hasan Khan, Arisa Himeno, Shouhei Kosugi, Yosuke Nakashima, Abdur Rafique, Ayana Imamura, Takaaki Hatanaka, Dai-Ichiro Kato, Yuji Ito
Chicken egg yolk immunoglobulin (IgY) is a functional substitute for mammalian IgG for antigen detection. Traditional IgY purification methods involve multi-step procedures resulting in low purity and recovery of IgY. In this study, we developed a simple IgY purification system using IgY-specific peptides identified by T7 phage display technology. From disulfide-constrained random peptide libraries constructed on a T7 phage, we identified three specific binding clones (Y4-4, Y5-14, and Y5-55) through repeated biopanning...
July 31, 2017: Journal of Peptide Science: An Official Publication of the European Peptide Society
https://www.readbyqxmd.com/read/28757359/identification-of-cyclic-peptides-for-facilitation-of-transcellular-transport-of-phages-across-intestinal-epithelium-in-vitro-and-in-vivo
#17
Shunsuke Yamaguchi, Shingo Ito, Mio Kurogi-Hirayama, Sumio Ohtsuki
Methodology to enhance intestinal absorption of macromolecular drugs is an important challenge for developing next-generation biomedicines. So far, various cationic cell-penetrating peptides have been reported to facilitate uptake of certain bioactive proteins. However, cyclic peptides might be better candidates, as they are more metabolically stable than linear peptides. Accordingly, we hypothesized that suitable cyclic peptides would promote the absorption of macromolecules across intestinal epithelium. To test this idea, we adopted Caco-2 cell permeability assay as an in vitro human intestinal absorption model, and M13 phage as a model of macromolecules...
July 27, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28756139/a-universal-phage-display-system-for-the-seamless-construction-of-fab-libraries
#18
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
#19
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/28744632/combinatorial-screening-of-peptides-specific-ligands-of-death-receptor-dr5
#20
V M Ukrainskaya, A V Stepanov, A A Belogurov, A G Gabibov
Death receptors, in particular DR5, are highly attractive targets of antitumor therapy. The major limitation to application of natural death receptor ligands (TRAIL) is their non-specific cytotoxicity against normal cells. Since TRAIL can also bind decoy receptors (DcR) and prevent induction of apoptosis, the search for new DR-specific ligands is a topical issue. In the present study, we used combinatorial phage display peptide libraries to select a panel of DR5-binding amino acid sequences. A comparative analysis of the selected peptides enabled identification of the consensus sequence responsible for binding to DR5...
July 25, 2017: Bulletin of Experimental Biology and Medicine
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