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Selection of Nanobodies that Block the Enzymatic and Cytotoxic Activities of the Binary Clostridium Difficile Toxin CDT
Abstract: The spore-forming gut bacterium Clostridium difficile is the leading cause of antibiotic-associated diarrhea in hospitalized patients. The major virulence factors are two large glucosylating cytotoxins. Hypervirulent strains (e.g. ribotype 027) with higher morbidity and mortality additionally produce the binary CDT toxin (Clostridium difficile transferase) that ADP-ribosylates actin and induces microtubule-based cell protrusions. Nanobodies are robust single domain antibodies derived from camelid heavy chain antibodies. Here we report the generation of functional nanobodies against the enzymatic CDTa and the heptameric receptor binding subunit CDTb. The nanobodies were obtained from a variable-domain repertoire library isolated from llamas immunized with recombinant CDTa or CDTb. Five CDTa-specific nanobodies blocked CDTa-mediated ADP-ribosylation of actin. Three CDTa-specific and two CDTb-specific nanobodies neutralized the cytotoxicity of CDTa+b. These nanobodies hold promise as new tools for research, diagnosis and therapy of C. difficile associated disease
Single-Domain Antibodies: Biology, Engineering and Emerging Applications
Single-domain antibodies (sdAbs) represent the minimal antigen binding-competent form of the immunoglobulin domain and have unique properties and applications. SdAbs are naturally produced as the variable domains of the heavy chain-only antibodies of camelid ruminants and cartilaginous fishes, but can also be engineered synthetically from autonomous human or mouse VH or VL domains. The scope of this research topic and associated e-book covers current understanding and new developments in (i) the biology, immunology and immunogenetics of sdAbs in camelids and cartilaginous fishes, (ii) strategies for sdAb discovery, (iii) protein engineering approaches to increase the solubility, stability and antigen-binding affinity of sdAbs and (iv) specialized applications of sdAbs in areas such diagnostics, imaging and therapeutics.
Biochemical Sensors (In 2 Volumes)
This book covers the full scope of biochemical sensors and offers a survey of the principles, design and applications of the most popular types of biosensing devices. It is presented in 19 chapters, written by 20 distinguished scientists as well as their co-workers. The topics include the design of signal transducers, signal tags and signal amplification strategies, the structure of biosensing interfaces with new biorecognition elements such as aptamers and DNAzymes, and different newly emerging nanomaterials such as Au nanoclusters, carbon nitride, silicon, upconversion nanoparticles and two-dimensional materials, and the applications in wearable detections, biofuel cells, biomarker analyses, bioimaging, single cell analysis and in vivo sensing.By discussing recent advances, it is hoped this book will bridge the common gap between research literature and standard textbooks. Research into biochemical sensors and their biomedical applications is proceeding in a number of exciting directions, as reflected by the content. This book is published in honor of the 90th birthday of Professor Shaojun Dong, who performed many pioneering studies on modified electrodes and biochemical sensors.
Single Domain Antibodies
The development of the hybridoma technology created the possibility to obtain unlimited amounts of monoclonal antibodies (mAb) with high specificity and affinity for any target and to introduce mAbs in a wide range of applications; however, the bulky size of mAbs, costly production, and cumbersome engineering hampered regularly their streamlined development in some applications. In Single Domain Antibodies: Methods and Protocols, expert researchers examine single variable domain antibody fragments, referred to as VH, VL, VHH or VNAR. These fragments are the smallest intact antigen-binding fragments that can be produced recombinantly at low cost. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Immunoinformatics
In contrast to existing books on immunoinformatics, this volume presents a cross-section of immunoinformatics research. The contributions highlight the interdisciplinary nature of the field and how collaborative efforts among bioinformaticians and bench scientists result in innovative strategies for understanding the immune system. Immunoinformatics is ideal for scientists and students in immunology, bioinformatics, microbiology, and many other disciplines.
Temporal and Spatial Regulation of Plant Genes
First attempts to isolate plant genes were for those genes that are abun dantly expressed in a particular plant organ at a specific stage of devel opment. However, many important gene products are produced in a very minute quantity and in specialized cell types. Such genes can now be isolated using a variety of approaches, some of which are described in this volume. The rapid progress during the last decade in regeneration of a number of crop plants and the availability of molecular tools to introduce foreign genes in plants is allowing the engineering of specific traits of agri cultural importance. These genes must, however, be regulated in a spatial and temporal manner in order to have des...
Higglety Pigglety Pop!
‘A daring imagination has woven a simple rhyme into a brilliantly original tale [about Jennie, the Sealyham terrier, who seeks Experience and becomes the star of the World Mother Goose Theatre].' 'H. ‘Superb fantasy.' 'BL. Notable Children's Books of 1967 (ALA) 1968 Fanfare Honor List (H) Best Books of 1967 (SLJ) Children's Books of 1967 (Library of Congress)
Islam Against the West
This book gives a unique perspective on the interwar history of the Middle East. By telling the life story of one man, it illuminates the political and cultural struggles of an era. Shakib Arslan (1869–1946) was a leading member of the generation of Ottoman Arabs who came to professional maturity just before the final defeat of the Ottoman Empire. Born to a powerful Lebanese Druze family, Arslan grew up perfectly suited to his time and place in history. He was one of the leading writers of his day and a dexterous, ambitious politician. But, by the end of World War I, Arslan and others of his generation found themselves adrift in a world no longer of their choosing, as the once great Ottoma...
A Field Guide to the Birds of South America
The only field guide to illustrate and describe every non-passerine species of bird in South America.
Los Que Mandan
This pioneering work, now available for the first time in English, seeks to analyze the political process in Argentina, a nation that has long aspired to political leadership of Latin America but has failed to fulfill its aspiration because of political fragmentation and factionalism. In determining who holds power in Argentina, Professor Imaz assembled information on the social backgrounds of political leaders and party workers, military officers, large landowners, managers and owners of industry and commerce, the hierarchy of the Roman Catholic Church, and officials of labor unions. This considerable amount of data for the years 1936–1961 provides the basis for a comparison of the processes of recruitment and the different social outlooks of the various elites. Professor Imaz's frequently cited book has gone through six printings in Argentina since first publication in 1964. For the English translation, he has added material that brings the data up to date. Professor Astiz has provided an introduction that places the study in context for those who are not familiar with Argentine history and politics.
