Immobilized Biomolecules In Analysis

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.

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.

Published since 1959, Advances in Applied Microbiology continues to be one of the most widely read and authoritative review sources in Microbiology.The series contains comprehensive reviews of the most current research in applied microbiology. Recent areas covered include bacterial diversity in the human gut, protozoan grazing of freshwater biofilms, metals in yeast fermentation processes and the interpretation of host-pathogen dialogue through microarrays.Impact factor for 2003: 1.893

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.

This volume presents discussions of theoretical and experimental considerations that have led to the analytical affinity chromatography field, as well as current efforts to use this methodology to characterize the interaction mechanisms of biological macromolecules and to establish conditions for employing bioaffinity chromatographic systems as preparative tools. The chapters include a comprehensive discussion of interactive chromatography theory (DeLisi and Hethcote), a review of experimental data obtained for biological macromolecules and the relevant theoretical considerations of affinity chromatography which led to them (Swaisgood and Chaiken), an evaluation of rate processes in affinity chromatography and the potential to determine biologically meaningful chemical rate constants (Walters), and the use of quantitative and molecular considerations to design affinity chromatographic systems (Stellwagen and Liu).

Handbook of Immunoassay Technologies: Approaches, Performances, and Applications unravels the role of immunoassays in the biochemical sciences. During the last four decades, a wide range of immunoassays has been developed, ranging from the conventional enzyme-linked immunosorbent assays, to the smartphone-based point-of-care formats. The advances in rapid biochemical procedures, novel biosensing schemes, fully integrated lab-on-a-chip platforms, prolonged biomolecular storage strategies, device miniaturization and interfacing, and emerging smart system technologies equipped with personalized mobile healthcare tools are paving the way to next-generation immunoassays, and are all discussed in this comprehensive text. Immunoassays play a prominent role in clinical diagnostics as they are the eyes of healthcare professionals, helping them make informed clinical decisions via confirmed disease diagnosis, and thus enabling favorable health outcomes. The faster and reliable diagnosis of infections will further control their spread to uninfected persons. Similarly, immunoassays play a prominent role in veterinary diagnostics, food analysis, environmental monitoring, defense and security, and other bioanalytical settings. Therefore, they enable the detection of a plethora of analytes, which includes disease biomarkers, pathogens, drug impurities, environmental contaminants, allergens, food adulterants, drugs of abuse and various biomolecules. Provides a valuable increase of understanding of cellular and biomedical functions Gives the most updated resource in the field of immunoassays, providing the comprehensive details of various types of immunoassays that need to be performed in healthcare, and in industrial, environmental and other biochemical settings Discusses all multifarious aspects of immunoassays Describes the immunoassay formats, along with their principle of operation, characteristics, pros and cons, and potential biochemical and bioanalytical applications Provides extensive knowledge and guided insights as detailed by experienced, renowned experts and key opinion makers in the field of immunoassays

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.

This book covers optical chemical sensing by means of optical waveguides, from the fundamentals to the most recent applications. The book includes a historical review of the development of these sensors, from the earliest laboratory prototypes to the first commercial instrumentations. The book reprints a lecture by the Nobel Laureate Charles Townes on the birth of maser and laser, which lucidly illustrates the development of new science and new technology.