Design, Selection, and Development of Novel Peptide Ligands for Bioseparations and Diagnostics PDF Download
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Author: Stefano Menegatti
Publisher:
ISBN:
Category :
Languages : en
Pages : 387
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Book Description
Author: Stefano Menegatti
Publisher:
ISBN:
Category :
Languages : en
Pages : 387
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Book Description
Author: Danielle Nicole Dremann
Publisher:
ISBN:
Category : Biochemistry
Languages : en
Pages : 275
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Book Description
ABSTRACT THE DEVELOPMENT OF PEPTIDE LIGANDS TO TARGET H69 by DANIELLE NICOLE DREMANN December 2015 Advisor: Prof. Christine S. Chow Major: Chemistry (Biochemistry) Degree: Doctor of Philosophy In the development of peptide ligands to target H69, SPPS and ESI MS was used to determine if 1) peptides could bind to modified H69 and 2) if increased affinity for the target RNA could be enhanced with modification. An alanine and arginine scan was synthesized and tested for this determination. Selected peptides were then tested using biophysical techniques such as circular dichroism and isothermal titration calorimetry. An assay was also designed to increase the efficiency of peptide ligand selection in which novel peptides with high affinity and selectivity could be identified for future projects. This assay was done using flow cytometry, instrumentation capable of identifying beads, bound to the target RNA conjugated to a fluorophore, based on fluorophore emission, and sorting them into 96-well plates for MS analysis. The last part of the research focused on aminoglycoside-H69 RNA interactions. ESI MS was used to obtain binding affinities and stoichiometries of 2AP- and A-containing H69 RNAs. The findings revealed that the binding mode had not changed between these two sets of RNAs, which revealed the value for using ESI MS in combination with other techniques, such as fluorescence, to give a complete picture of the binding mode (stoichiometry, affinity, selectivity) in comparison with conformational changes that may occur upon binding. Further exploration of aminoglycoside-H69 RNA interactions took place with the H69 peptide NQVANHQ-NH2 to determine whether a fragment-based drug design approach could be used to create small compounds for future in vivo applications.
Author: Nika Kruljec
Publisher:
ISBN:
Category :
Languages : en
Pages : 133
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Book Description
Author: Sophia Alexandra Kenrick
Publisher:
ISBN: 9781109483222
Category :
Languages : en
Pages : 294
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Book Description
Analysis of bacterial display libraries by flow cytometry enabled direct measurements of apparent equilibrium affinity, specificity, and kinetic dissociation rates. However, avidity effects due to high peptide display level and complex interactions between the target protein and bacterial cell surface masked themselves as both affinity and specificity, necessitating the use of cell-free measurements to corroborate affinity rankings made on the cell surface. Both mathematical modeling and experimental analysis of these effects were performed with multimeric protein targets--VEGF and streptavidin (SA)--to show how high peptide display level can mimic high peptide affinity. Further analysis of protein-peptide interactions using surface plasmon resonance (SPR) elucidated differences among affinity measurements with peptides expressed on the cell surface, peptides expressed as fusions to larger carrier proteins, and synthetically prepared peptides. These studies will facilitate future identification, analysis, and improvement of bioactive peptides for diagnostics and therapeutics.
Author: Amanda Jean Hughes Gilliam
Publisher:
ISBN: 9781339125404
Category :
Languages : en
Pages :
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Book Description
The caveolins are a family of human membrane proteins implicated in cancer and other diseases. In Chapter 1, I briefly explain some of the basic biochemistry of the caveolins, and why they are not only a promising target for research and therapeutics but also a particularly challenging target. The novel peptide WL47, the development of which is described in later chapters, is introduced as a caveolin-1 ligand. WL47 will allow new avenues of investigation for this challenging target. In Chapter 2, I review, compare, and contrast some of the myriad techniques available to quantify binding affinity for high affinity binding interactions such as the interaction between caveolin-1 and WL47. As I discovered in my own attempts to measure the dissociation constant for this interaction, high affinity is a boon for future use of a ligand probe, but it can be challenging to quantify initially. Because a review of techniques specifically suited to high affinity measurement was not available at the time I needed it, I have written this chapter to fill this gap in the literature. In Chapter 3, I discuss the development of peptide ligand WL47 through an iterative process of library design, synthesis, screening, and data analysis. High affinity binding, target selectivity, and activity as an inducer of deoligomerization of oligomeric caveolin-1 is presented. Beyond providing a novel tool for probing the function of caveolin-1 oligomerization, this chapter is also a roadmap for the creation of future peptide ligands through this generalizable, iterative process. Chapter 4 details the progress that has been made toward understanding the importance of WL47 dimerization through a disulfide bridge. This chapter also covers initial efforts towards designing a redox-stable version of WL47 that will remain functional in conditions like those found in the interior of a live cell. Chapter 5 concludes the dissertation with a discussion of the current state of the caveolin ligand project, including detailed descriptions of the experiments that will be necessary for future researchers to bring this project to its conclusion.
Author: Greg T. Hermanson
Publisher: Academic Press
ISBN:
Category : Science
Languages : en
Pages : 494
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Book Description
This book is a practical guide to the preparation and use of immobilized affinity ligands for purification, catalysis, and analysis. Special emphasis is given to immunochemical techniques including antibody isolation, preparation of antibody fragments using immobilized enzymes, and immunoaffinity chromatography. The book provides easy-to-follow, well-tested protocols to allow the uninitiated to use these techniques to the maximum advantage with minimum hassle. In addition, it shows researchers how to save money by making their own optimized affinity supports. Matrix activation: Ligand immobilization, Binding and elution of target molecules, Enzyme catalysis on solid supports, Analytical affinity chromatography, Isolation/purification of antibodies, Preparation of antibody fragments, Immunoaffinity chromatography, Immobilization of nucleic acids, Use of immobilized ligands for removal of trace contaminants Practical advice on choosing: Matrices, Spacers, Methods of activation and coupling Background information and insights on: Affinity interactions, The ease and power of affinity chromatography, Attaching molecules to insoluble supports, Matrices currently in use, Over 20 methods of activation, Spacers, Extensive References
Author: Roger G. Harrison
Publisher: Oxford University Press
ISBN: 0190213736
Category : Technology & Engineering
Languages : en
Pages : 577
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Book Description
Designed for undergraduates, graduate students, and industry practitioners, Bioseparations Science and Engineering fills a critical need in the field of bioseparations. Current, comprehensive, and concise, it covers bioseparations unit operations in unprecedented depth. In each of the chapters, the authors use a consistent method of explaining unit operations, starting with a qualitative description noting the significance and general application of the unit operation. They then illustrate the scientific application of the operation, develop the required mathematical theory, and finally, describe the applications of the theory in engineering practice, with an emphasis on design and scaleup. Unique to this text is a chapter dedicated to bioseparations process design and economics, in which a process simular, SuperPro Designer® is used to analyze and evaluate the production of three important biological products. New to this second edition are updated discussions of moment analysis, computer simulation, membrane chromatography, and evaporation, among others, as well as revised problem sets. Unique features include basic information about bioproducts and engineering analysis and a chapter with bioseparations laboratory exercises. Bioseparations Science and Engineering is ideal for students and professionals working in or studying bioseparations, and is the premier text in the field.
Author: Alexander von Gabain
Publisher: Springer Science & Business Media
ISBN: 3709107091
Category : Medical
Languages : en
Pages : 318
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Book Description
“Development of novel vaccines” gives an overview of the tasks in basic research leading to the final product – the vaccine and its applications, belonging to the most complex biologics in the pharmaceutical field. Distinct from most textbooks in the vaccine arena, the current issue focuses on the translational aspect, namely, how research results can be transformed into life-saving medical interventions. Each chapter of the book deals with one important paradigm for the development of novel vaccines, along the value chain towards the final vaccine, and furthermore, with the inevitable tools required for this process. Contributions are prepared by teams of scientists, all of whom are experts in the field, most of them anchored in biomedical organizations devoted to translational culture, thereby lighting the certain topics from different views. This volume is a must read for researchers engaged in vaccine development and who really want to see their research results to become a product.
Author: Heinrich Klefenz
Publisher: Wiley-VCH
ISBN:
Category : Medical
Languages : de
Pages : 328
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Book Description
This volume focuses on pharmaceutical biotechnology as a key area of life sciences. The complete range of concepts, processes and technologies of biotechnology is applied in modern industrial pharmaceutical research, development and production. The results of genome sequencing and studies of biological-genetic function are combined with chemical, micro-electronic and microsystem technology to produce medical devices and diagnostic biochips. A multitude of biologically active molecules is expanded by additional novel structures created with newly arranged gene clusters and bio-catalytic chemical processes. New organisational structures in the co-operation of institutes, companies and networks enable faster knowledge and product development and immediate application of the results of research and process development. This book is the ideal source of information for scientists and engineers in research and development, for decision-makers in biotech, pharma and chemical corporations, as well as for research institutes, but also for founders of biotech companies and people working for venture capital corporations.
Author: National Research Council
Publisher: National Academies Press
ISBN: 0309047854
Category : Science
Languages : en
Pages : 133
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Book Description
The ability of the United States to sustain a dominant global position in biotechnology lies in maintaining its primacy in basic life-science research and developing a strong resource base for bioprocess engineering and bioproduct manufacturing. This book examines the status of bioprocessing and biotechnology in the United States; current bioprocess technology, products, and opportunities; and challenges of the future and what must be done to meet those challenges. It gives recommendations for action to provide suitable incentives to establish a national program in bioprocess-engineering research, development, education, and technology transfer.
