Immobilized Biomolecules In Analysis
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Immobilized Biomolecules In Analysis
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Author : Tony Cass
language : en
Publisher: Oxford University Press
Release Date : 1999-01-28
Immobilized Biomolecules In Analysis written by Tony Cass and has been published by Oxford University Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 1999-01-28 with Science categories.
Biomolecules and cells are critical components of biosensors and biomaterials, but in order to function in an artificial environment, they must be immobilized in a manner that does not affect their interaction with target analytes. Biosensors demonstrate that we can harness the incredible functions of living molecules and cells for our own purposes and are therefore at the forefront of technology. Moreover the applications of immobilized biomolecules and cells are expected to expand far beyond biosensor applications and indeed are already used for pharmaceutical production and testing. Biomaterials will become increasing common as they are being developed into toxic filters, artificial organs, and even silicon chips. This book provides a selection of methods for the immobilization of biomolecules and cells on a variety of surface with different geometries and chemistries so that they retain their function and guidelines on which method to use. Also included are the analytical techniques to measure the functionality of immobilized biomolecules. All the protocols have been tried and validated by the authors. Immobilized Biomolecules in Analysis: A Practical Approach is an invaluable guide to all researchers in the fields of biosensors and biomaterials. Research in biosensors is carried out in a wide variety of fields including biochemistry, chemistry, engineering, laboratory medicine, environmental and defence research. The protocols are written so that an extensive prior knowledge of biochemistry is not required to use them.
Development Of Methods For Directed And Structured Immobilization Of Biomolecules
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Author : Björn Waterkotte
language : en
Publisher: Cuvillier Verlag
Release Date : 2013-11-07
Development Of Methods For Directed And Structured Immobilization Of Biomolecules written by Björn Waterkotte and has been published by Cuvillier Verlag this book supported file pdf, txt, epub, kindle and other format this book has been release on 2013-11-07 with Science categories.
Surface immobilized molecules play a crucial role in applied as well as in basic research. They can be found in DNA or protein-array as well as in cell-migration or biochemical interaction experiments. Especially for proteins the immobilization can critically affect molecule accessibility and activity. In this work two essential aspects of protein immobilization were investigated: the identification of suitable surface functionalization by two array approaches and the generation of laterally structured gray scale protein patterns by projection lithography. In proof-of-concept experiments a new strategy to create an array of different, neighboring functionalizations on the same surface was successfully tested. In "mechanical protection" inert plungers were applied to protect distinct surface areas from the surface functionalization reagents and allowed a selective modification of individual regions in serial reactions. In a particle based alternative approach a high spot density surface array was fabricated using functionalized, polyvinyl acetate, melamine and silicon oxide-based particles. The in situ encoded bead-based array (IEBA) allowed the study of immobilization conditions and the effect of protein adhesion. For the reliable and fast analysis of array-images with several thousand individual particles a custom open-source evaluation software was developed. To create protein patterns a custom-built projection lithography device was utilized to generate high definition protein patterns. Patterning conditions were optimized with respect to deposition speed and signal to noise ratio while retaining a biocompatible process flow. The established technology provides protein patterns with great control over the deposited surface density, patterns were transferred within seconds to minutes to functionalized glass slides. To obtain three-dimensional, surface-functionalized surfaces this approach was transferred to thin polymer films. The substrates were demonstrated to be biocompatible with mouse fibroblasts. These surfaces allow a novel level of control addressing both the chemical surrounding and the shape of a cell microenvironment.
A Chemical Investigation Of Immobilized Biological Molecules In Clinical Analysis
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Author : Gary Lee Smith
language : en
Publisher:
Release Date : 1977
A Chemical Investigation Of Immobilized Biological Molecules In Clinical Analysis written by Gary Lee Smith and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1977 with Binding sites (Biochemistry) categories.
Analytical Applications Of Immobilized Enzyme Reactors
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Author : S. Lam
language : en
Publisher: Springer
Release Date : 1994
Analytical Applications Of Immobilized Enzyme Reactors written by S. Lam and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 1994 with Medical categories.
Enzymes are bio-catalysts which effect transformation of substrates to products with high specificity. The usage of enzymes in domestic and industrial applications is well known and has been well documented since the early history of civilization. With the advances in understanding of enzymology, usage of enzymes in industrial and biotechnological pro cesses and molecular medicine has proliferated. One of the key factors in the widespread application of enzymes in modern technologies is the development of enzyme immobilization tech niques, which overcome certain practical, functional and economic con straints. Many natural enzymes can be stabilized by immobilization on solid matrices, with most of the activity retained, for a variety of applica tions. An important application of immobilized enzymes is in liquid chromatography. In the last decade, post-column enzyme detection has become established as an important discipline in liquid chromatography. The new detection approach offers more sensitive and specific ways for measuring major classes of biomolecules. Reactors are fabricated by packing the immobilized enzymes into small columns, which can be placed immediately after an HPLC column.
Uses Of Immobilized Biological Compounds
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Author : George G. Guilbault
language : en
Publisher: Springer Science & Business Media
Release Date : 2012-12-06
Uses Of Immobilized Biological Compounds written by George G. Guilbault and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-06 with Technology & Engineering categories.
In Uses of Immobilized Biological Compounds the reader will find a comprehensive survey of the field written by acknowledged experts who met in Brixen, Italy, between May 9 and 14, 1993 for a NATO Advanced Research Workshop devoted to the topic. The resulting volume presents a critical review of the latest results in the area and sets guidelines for future research. The 53 reports presented here cover: (A) General Aspects of Immobilizing Biological Compounds; (B) Medical, Clinical and Pharmaceutical Applications; (C) Electrochemical Biosensors; (E) Defense Applications; (F) Immunosensors and Receptors; (G) Food, Environmental, Clinical and Analytical Applications; and (H) Biotechnology and Marketing. In short, all aspects of the area are presented, in a compact format which will appeal to undergraduates, technicians, and professional scientists in the food, clinical, environmental, pharmaceutical and industrial fields.
Immobilization Free Electrochemical Dna Based Bio Analysis
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Author : Xiaoteng Luo
language : en
Publisher:
Release Date : 2011
Immobilization Free Electrochemical Dna Based Bio Analysis written by Xiaoteng Luo and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2011 with Biomolecules categories.
Real Time Analysis Of Biomolecular Interactions
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Author : K. Nagata
language : en
Publisher: Springer Science & Business Media
Release Date : 2013-12-11
Real Time Analysis Of Biomolecular Interactions written by K. Nagata and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2013-12-11 with Science categories.
The metabolism of all living organisms consists of elaborate and complex interactions among numbers of biomolecules, including protein-protein interactions. BIACORE is a new device that uses surface plasmon resonance for kinetic analysis of molecular interactions and automates whole analytical processes. The number of users of BIACORE is increasing worldwide, but until now nothing had been published in English to explain the principles of this new methodology and the kinds of applications it makes available. This updated and revised version of the Japanese original explains the underlying principles of BIACORE, providing concrete examples that utilize the methodology for analysis in areas ranging from basic to applied sciences. With its generous use of illustrations, this book is a valuable source of information for all users of BIACORE.
Investigation Of The Interactions Between Biomolecules And Mesoporous Inorganic Materials In Biomolecule Immobilization For Bioseparation And Biocatalysis
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Author : Jungseung Kim
language : en
Publisher:
Release Date : 2011
Investigation Of The Interactions Between Biomolecules And Mesoporous Inorganic Materials In Biomolecule Immobilization For Bioseparation And Biocatalysis written by Jungseung Kim and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2011 with categories.
The interactions between biomolecules and solid surfaces are complex phenomena. Understanding the nature of these interactions can allow engineering highly efficient systems for bioseparation and biocatalysis. However, there is still a lack of understanding of these fundamental interactions due to the complexity and fragility of biomolecules, especially proteins. The overall goal in this research is to improve the current understanding of these interactions as functions of the properties of mobile phases and stationary phases by investigating adsorption isotherms, adsorption thermodynamics, and biocatalytic activity of immobilized proteins. Mesoporous silica and alumina were used as stationary phases (adsorbents). In particular, mesostructured cellular foam (MCF) silica, which has an open 3-dimensional pore structure with superior physical properties, was used to immobilize biomolecules. The surface chemistry of the synthesized MCF silica was engineered to control the immobilization of biomolecules by grafting functional groups, including charge-terminated (amine-terminated, mecapto-terminated) and hydrophobic-terminated groups (methyl-terminated) groups, to the surface. The interactions between biomolecules and prepared adsorbents were also investigated using flow microcalorimetry (FMC) to reveal the adsorption mechanisms during the immobilization of biomolecules at different levels of pH and ionic strength and several functionalized solid surfaces. Adsorption thermodynamics and mechanism can be modulated by changing ionic strength by adding a neutral salt (sodium sulfate) and by changing the pH. Biomolecule adsorption is a complex phenomenon, exhibiting multiple heat events. However, similar thermograms were observed for the interactions between protein and MCF silicas in most cases. Also, the driving force for protein adsorption was investigated by using semi-empirical analysis. Adsorption energetics were affected significantly by surface modification. Also, the FMC data along with batch adsorption isotherm revealed that aminopropyl-grafted MCF silica has the property of ion-exchanger. The energetics of different biomolecules (tryptophan, lysozyme, and bovine serum albumin) was investigated in different pH. The number of distinct exothermic peaks corresponded to the number of binding sites of biomolecules. And the magnitude of net heat of adsorption was increased according to increasing molecular weight at pH 5.2. However, the energetics of bovine serum albumin was significantly changed from pH 5.2 to pH4. It is attributed to the conformational change of protein as the function of pH. The biocatalytic activity of immobilized enzyme (lipase from Pseudomonas fluorescens) was investigated on different functionalized MCF silicas to understand the effect of surface modification. The amine-terminated long functional group showed the most positive effect to increase the catalytic efficiency (kcat/Km) of immobilized enzyme due to the orientation change and the mixed interactions (charged interactions and hydrophobic) interactions. The biocatalytic activity of immobilized enzyme (lipase from Pseudomonas cepacia) was further extended by using functionalized aluminas as enzyme supporting materials. The measurement of the biocatalytic activity of immobilized enzymes showed that the effect of surface modification has important roles for the activation of immobilized enzyme activity and the optimal enzyme supporting material need to be selected. This study provides insights into the natures and mechanisms of biomolecule immobilization on mesoporous silica to design highly efficient systems for biomolecule immobilization for bioseparation and immobilized enzyme technology for biocatalysis.
Immobilized Cells And Organelles
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Author : Mattiasson
language : en
Publisher: CRC Press
Release Date : 2018-01-18
Immobilized Cells And Organelles written by Mattiasson and has been published by CRC Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018-01-18 with Science categories.
Cells and organelles are small units for biochemical synthetic purposes, often the smallest practically feasible unit since they contain coenzyme regenerating system, ordered enzyme sequences, etc.These volumes, besides giving some insight into basic technology (immobilisation procedures, etc.), also sum up the current know-how in this subject area and try to predict some future trends.The termimmobilized cells covers everything from dead cells with a single active enzyme species to cells proliferating on or within a three dimensional polymer matrix. The practical handling of these structures make them useful in various applications, e.g. large-scale production of biomolecules, biodegration, analysis, etc.
Surface Modifications For Enhanced Immobilization Of Biomolecules
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Author : Yunling Bai
language : en
Publisher:
Release Date : 2006
Surface Modifications For Enhanced Immobilization Of Biomolecules written by Yunling Bai and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2006 with Biochips categories.
Abstract: The goal of this study is to investigate the application of immobilization technology in various systems: immobilized cell/enzyme bioreactors, affinity chromatography, and BioMEM surface modification. These systems were investigated to solve a particular problem. A novel method for the co-immobilization of whole cells and LDH enzyme on cotton cloth was developed using poly (ethyleneimine) (PEI), which induced the formation of PEI-enzyme-cell aggregates and their adsorption onto cotton cloth, leading to multilayer co-immobilization of cells and enzyme with a high loading amount (0.5 g cell and 8 mg LDH per gram of cotton cloth) and activity yield (>95%). A fibrous bed bioreactor with cells and enzyme co-immobilized on the cotton cloth was then evaluated for R-HPBA production in fed-batch and repeated batch modes, which gave relatively stable reactor productivity. A novel surface treatment method using poly(ethyleneimine) (PEI), an amine-bearing polymer, was developed to enhance antibody binding on the poly(methyl methacrylate) (PMMA) microfluidic immunoassay device. By treating the PMMA surface of the microchannel on the microfluidic device with PEI, 10 times more active antibodies can be bound to the microchannel surface as compared to those without treatment or treated with the small amine-bearing molecule, hexamethylene diamine (HMD). Consequently, PEI surface modification greatly improved the immunoassay performance of the microfluidic device, making it more sensitive and reliable in the detection of IgG. The surface modification method was further simplified and optimized to enhance polymer-based microchannel ELISA for E. coli O157:H7 detection. By applying an amine-bearing polymer, poly (ethyleneimine) (PEI), onto a poly (methyl methacrylate) (PMMA) surface at pH higher than 11, PEI molecules were covalently attached and their amine groups were introduced to the PMMA surface. Zeta potential analysis and X-ray photoelectron spectroscopy (XPS) demonstrated that the alkaline condition is preferable for PEI attachment onto the PMMA surface. Compared to untreated PMMA microchannels, ~45 times higher signal and 3 times higher signal/noise ratio were achieved with the PEI surface treatment, which also shortened the time required for cells to bind to the microchannel surface to ~2 minutes, much less than that usually required for the same ELISA carried out in 96-well plates.