Download or read book Method Optimization of Aqueous Size Exclusion Chromatography for Six Proteins a Study of Lysozyme Ovalbumin Bovine Serum Albumin Human Serum Albumin Immunoglobulin and Thyroglobulin by Size Exclusion Chromatography Including Degradation Studies written by Elizabeth Davidson and published by . This book was released on 2021 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: "High-performance liquid chromatography (HPLC) is used often for the separation of proteins or other biomolecules. One type of HPLC used to determine the purity of a protein is size-exclusion chromatography (SEC). The size-based separation in SEC occurs when smaller molecules are retained in the pores of the stationary phase, while larger molecules are eluted first. This allows aggregates of proteins as well as any small contaminants to be clearly separated. SEC can be used either with aqueous or organic mobile phase. In this study, six different proteins were studied using aqueous SEC: bovine serum albumin, human serum albumin, immunoglobulin, lysozyme, ovalbumin, and thyroglobulin. Mobile phase conditions were optimized for separating the monomer from the aggregates of each of these compounds. A calibration curve was constructed using the monomer of each of the proteins. The proteins were then degraded under different conditions, including acid, base, oxidation, and UV light to study the molecular weight effects of degradation on proteins. The method was then evaluated using various validation studies. It was found that separating similarly sized proteins is very difficult in SEC, but that the aggregates of each protein can be separated well from the monomer."--

Download or read book Development and Validation of an Analytical Method for the Separation of a Mixture of Human Serum Albumin Ovalbumin and Immunoglobulin Including Their Forced Degradation Studies Using Size exclusion Chromatography written by Nishal Patel and published by . This book was released on 2021 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: "HSA is a protein in blood with a molecular mass 66.5 k Da. Ovalbumin is the main protein found in egg white with a molecular mass 45 k Da. Immunoglobulin – It is an antibody, large Y-shaped protein used by immune system & neutralize foreign objects such as pathogenic bacteria and viruses with a molecular mass 150k Da.A method was developed for the separation of a group of proteins (HSA, OVA, IgG) using Size Exclusion Chromatography. The method was developed on the Agilent HPLC system 1260 series with Diode Array Detection. Desired separation was achieved on Agilent, Advance Bio SEC, 300 A°, 2.7 μm, 4.6x300 mm (p/N) 1580-5301 column with isocratic mode by using 25 mM Potassium Phosphate buffer, pH-7.0 and 100 mM NaCl as mobile phase. The Analysis was carried out using flow rate- 0.35 ml/ min, detection wavelength –220nm, column temperature -30°C and Injection Volume 3μl. The developed method was successfully applied for the separation of mixture of proteins, and it was validated per ICH guidelines in term of linearity, Precision, Robustness, specificity, System suitability and Stability."--

Download or read book Method Development and Validation for the Separation of Chymotrypsin Bovine Serum Albumin Ovalbumin and Lysozyme Using Reversed Phase Chromatography Mass Spectrometry written by Sussan Oladipo and published by . This book was released on 2024 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: "Reversed-phase liquid chromatography (RPLC) has been demonstrated to be a promising separation technique in separating proteins and peptides. The efficiency obtained with the RPLC is generally superior to other separation techniques such as size-exclusion chromatography, ion-exchange chromatography, hydrophilic interaction chromatography, and hydrophobic interaction chromatography. Another advantage of RPLC is its ability to couple with mass spectrophotometry detection and use in forced-degradation studies. A simple reversed-phase liquid chromatographic method was developed to separate a sample of chymotrypsin, lysozyme, bovine serum albumin, and ovalbumin. The method was developed on a 25 mm Phenomenex Column C8, Particle size: 5 μm, I.D. 4.6 mm under 50oC column temperature. Mobile phase used consisted of 1.0% trifluoroacetic acid in deionized water and Acetonitrile at a flow rate of 1.0 ml/min with UV-Vis detection at 210, 220 and 280 nm using DAD detector. The separation was conducted under gradient elution with 30 to 70% ACN gradient rate for 20 min. Forced Degradation studies was done on the BSA sample with resulting significant degradation effect from acid, basic, hydrogen peroxide, UV Light and Heat incubations stress test on the BSA sample. Mass spectroscopic analysis of the degradants identifies amino acid fragments. The developed method was validated for robustness, linearity, accuracy, precision, detection limit and quantitation to quantify bovine serum albumin. The method proved simple, accurate and precise with over 97% average recovery of bovine serum albumin."--

Download or read book Modern Size Exclusion Liquid Chromatography written by Andre Striegel and published by John Wiley & Sons. This book was released on 2009-07-31 with total page 512 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Second Edition of Modern Size-Exclusion Chromatography offers a complete guide to the theories, methods, and applications of size-exclusion chromatography. It provides an unparalleled, integrated, up-to-date treatment of gel permeation and gel filtration chromatography. With its detailed descriptions of techniques, data handling, compilations of information on columns and column packings, and tables of important solvents and reference materials, the book offers readers everything they need to take full advantage of this popular macromolecular characterization technique. Since publication of the first edition in 1979, there have been many important advances in the field of size-exclusion chromatography. This Second Edition brings the book thoroughly up to date, with expert coverage of: New and emerging industrial and research applications Practical aspects of size-exclusion chromatography (SEC) and multidetector and multidimensional SEC technologies for polymer architecture and copolymer analysis Updated information on the latest equipment and techniques New best practices for the lab SEC in relation to polymer characterization techniques such as GPEC, LCCC, and rheology Throughout the text, detailed examples guide you step by step through all the latest techniques and applications. With its extensive revisions and updates written by leading experts and pioneers in the field, Modern Size-Exclusion Liquid Chromatography remains the definitive resource for the broad range of researchers and scientists who use HPLC and GPC methods.

Download or read book Protein Analysis and Purification written by I.M. Rosenberg and published by Springer Science & Business Media. This book was released on 2013-03-14 with total page 453 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.

Download or read book Protein Purification written by Jan-Christer Janson and published by John Wiley & Sons. This book was released on 1998 with total page 712 pages. Available in PDF, EPUB and Kindle. Book excerpt: This is a state-of-the-art sourcebook on modern high-resolution biochemical separation techniques for proteins. It contains all the basic theory and principles used in protein chromatography and electrophoresis.

Download or read book Bioconjugate Techniques written by Greg T. Hermanson and published by Academic Press. This book was released on 2010-07-26 with total page 1233 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioconjugate Techniques, 2nd Edition, is the essential guide to the modification and cross linking of biomolecules for use in research, diagnostics, and therapeutics. It provides highly detailed information on the chemistry, reagent systems, and practical applications for creating labeled or conjugate molecules. It also describes dozens of reactions with details on hundreds of commercially available reagents and the use of these reagents for modifying or cross linking peptides and proteins, sugars and polysaccharides, nucleic acids and oligonucleotides, lipids, and synthetic polymers. A one-stop source for proven methods and protocols for synthesizing bioconjugates in the lab Step-by-step presentation makes the book an ideal source for researchers who are less familiar with the synthesis of bioconjugates More than 600 figures that visually describe the complex reactions associated with the synthesis of bioconjugates Includes entirely new chapters on the latest areas in the field of bioconjugation as follows: Microparticles and nanoparticlesSilane coupling agentsDendrimers and dendronsChemoselective ligationQuantum dotsLanthanide chelatesCyanine dyesDiscrete PEG compoundsBuckyballs,fullerenes, and carbon nanotubesMass tags and isotope tagsBioconjugation in the study of protein interactions

Download or read book Protein Chromatography written by Giorgio Carta and published by John Wiley & Sons. This book was released on 2020-06-02 with total page 440 pages. Available in PDF, EPUB and Kindle. Book excerpt: An all-in-one practical guide on how to efficiently use chromatographic separation methods Based on a training course that teaches the theoretical as well as practical aspects of protein bioseparation to bioprocess professionals, this fully updated and revised new edition offers comprehensive coverage of continuous chromatography and provides readers with many relevant examples from the biopharmaceutical industry. Divided into two large parts, Protein Chromatography: Process Development and Scale-Up, Second Edition presents all the necessary knowledge for effective process development in chromatographic bioseparation, both on small and large scale. The first part introduces chromatographic theory, including process design principles, to enable the reader to rationalize the set-up of a bioseparation process. The second part illustrates by way of case studies and sample protocols how the theory learned in the first part may be applied to real-life problems. Chapters look at: Downstream Processing of Biotechnology Products; Chromatography Media; Laboratory and Process Columns and Equipment; Adsorption Equilibrium; Rate Processes; and Dynamics of Chromatography Columns. The book closes with chapters on: Effects of Dispersion and Rate Processes on Column Performance; Gradient Elution Chromatography; and Chromatographic Column Design and Optimization. -Presents the most pertinent examples from the biopharmaceutical industry, including monoclonal antibodies -Provides an overview of the field along with design tools and examples illustrating the advantages of continuous processing in biopharmaceutical productions -Focuses on process development and large-scale bioseparation tasks, making it an ideal guide for the professional bioengineer in the biotech and pharma industries -Offers field-tested information based on decades of training courses for biotech and chemical engineers in Europe and the U.S. Protein Chromatography: Process Development and Scale-Up, Second Edition will appeal to biotechnologists, analytical chemists, chromatographers, chemical engineers, pharmaceutical industry, biotechnological industry, and biochemists.

Download or read book Drug Discovery and Evaluation Pharmacological Assays written by Hans Vogel and published by Springer Science & Business Media. This book was released on 2007-10-30 with total page 2118 pages. Available in PDF, EPUB and Kindle. Book excerpt: The new edition of this successful reference offers both cutting-edge and classic pharmacological methods. Thoroughly revised and expanded to two volumes, it offers an updated selection of the most frequently used assays for reliably detecting the pharmacological effects of potential drugs. Every chapter has been updated, and numerous assays have been added. Each of the more than 1,000 assays comprises a detailed protocol outlining purpose and rationale, and a critical assessment of the results and their pharmacological and clinical relevance.

Download or read book Amino Acid Homopolymers Occurring in Nature written by Yoshimitsu Hamano and published by Springer Science & Business Media. This book was released on 2010-07-30 with total page 175 pages. Available in PDF, EPUB and Kindle. Book excerpt: Microorganisms are capable of producing a wide variety of biopolymers. Homopolymer peptides, which are made up of only a single type of amino acid, are far less ubiquitous. The only two amino-acid homopolymers known to occur in nature are presented in this volume. Poly-epsilon-L-lysine is a polycationic peptide and exhibits antimicrobial activity against a wide spectrum of microorganisms. It is both safe and biodegradable and is therefore used as a food preservative in several countries. In addition, there has been great interest in medical and other applications of poly-lysine and its derivatives. In contrast, poly-gamma-glutamic acid is an unusual anionic polypeptide. It is also water soluble, biodegradable, edible, non-toxic and non-immunogenic and can be chemically modified to introduce various drugs. These features are very useful for pharmaceutical and biomedical applications. Poly-glutamic acid is also a highly attractive as a food ingredient.

Download or read book Modern Methods in Analytical Morphology written by J. Gu and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 441 pages. Available in PDF, EPUB and Kindle. Book excerpt: While advances in modem medicine largely parallel our understanding of morphology, discoveries in morphology are propelled by developments of new tools and means to visualize and measure tissue elements. The invention of dissecting, light, fluorescence and electron microscopes together with advances in labeling and staining techniques are among the stepping stones of morphological progress. Today, we are in an exciting new era when classical morphology is being combined with developments from other disciplines. The combination of morphology and immunology resulted in immunocytochemistry; morphology and molecular biology led to in situ hybridization and in situ PCR. Adding computer science to morphology gave birth to image analysis. Combining laser technology and the microsope evolved into confocal microscope. For more than a decade, modem morphology has continued to develop by merging with other disciplines at a rate that is still gathering momentum, providing exciting and dynamic new frontiers for other biological fields. "Modem Methods in Analytical Morphology," based largely on the "First International Workshop on Modem Methods in Analytical Histochemistry, "is an updated review of the current trends in the field. It covers an extensive array of new technical developments in major disciplines of modem morphology. The authors are not only leaders in their fields but also have extensive "hands on" experience with "bench work. " Their chapters are written in a comprehensive manner including discussion of both theoretical considerations and practical applications to give the readers a broad view of the topics covered.

Download or read book Essential Immunology written by Ivan Maurice Roitt and published by . This book was released on 1971 with total page 244 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Chemicals via Higher Plant Bioengineering written by Fereidoon Shahidi and published by Springer Science & Business Media. This book was released on 1999-05-31 with total page 296 pages. Available in PDF, EPUB and Kindle. Book excerpt: Food and raw material for its production was generally produced via the traditional agriculture. On the other hand, novel chemicals were manufactured in the laboratory or extracted from plant and animal sources. However, as the world population is steadily in creasing, there is a decrease in traditional agriculture productivity and concerns are also expressed over the damage inflicted to the environment and restrictions that might be en forced in food production. At the same time, there is an increasing demand for high qual ity agricultural products as well as for food ingredients related to both the traditional or newly discovered nutrients or phytochemicals. Trends and developments,~n the area of plant biotechnology and bioengineering has allowed manipulation of genes' !lnd/or insertion of new genes, thus production of trans genic plants. Starting from the introduction of agronomic traits, particularly stress resis tance to diverse environmental factors, process and sensory characteristics, food quality and production of novel varieties of plant-based products through genetic engineering, biotechnology is changing the,;agriculture and the concept of production of plant-ba~~d raw materials. Increasing attention is being paid on research for production of plants !pat can provide a wide array of food and non-food products. Perhaps the first non-food pro,d uct that plant biotechnology would achieve is production of large scale custom-designed industrial oils, but the list of chemicals is long, ranging" from oils and specific triacyl glycerols to biopolymers, enzymes, blood components, amo~g others.

Download or read book Basic Protein and Peptide Protocols written by John M. Walker and published by Humana. This book was released on 1994-04-26 with total page 512 pages. Available in PDF, EPUB and Kindle. Book excerpt: Basic Protein and Peptide Protocols offers an excellent collection of reproducible, step-by-step laboratory methods covering three major areas: (1) the quantitation and characterization of proteins, (2) the electrophoretic and blotting procedures used in protein isolation and characterization, and (3) the analysis of protein and peptide structure. THOUSANDS of labs are already using Basic Protein and Peptide Protocols-you should be too!

Download or read book Practical HPLC Methodology and Applications written by Brian A. Bidlingmeyer and published by John Wiley & Sons. This book was released on 1993-05-06 with total page 470 pages. Available in PDF, EPUB and Kindle. Book excerpt: Of related interest. Trace and Ultratrace Analysis by HPLC Satinder Ahuja Written by a leading scientist in the field, this monograph provides the first definitive and technically up-to-date treatment of the theory, equipment, and applications of chemistry's most powerful reliable analytical technique. Coverage includes an encyclopedic compendium of common substances that require trace and ultratrace analysis, and features clear discussion of such important topics as considerations for HPLC equipment, sensitive detectors, sample preparation, method development, selectivity and computer-based optimizations, optimizing detectability, and much more. 1991 (0 471-51419-5) 432 pp. High Performance Liquid Chromatography in Biotechnology Edited by William S. Hancock Analytical chemists, biochemists, and chemical engineers will find this up-to-date guide to HPLC's recent developments essential for enhancing on-the-job technical expertise. Extensive coverage includes the broad applications of HPLC, ranging from major chromatographic techniques (including reversed phase, ion exchange, affinity and hydrophobic interaction chromatography) to specific separations such as those in monoclonal antibody and nucleic acid purification. Techniques for quality control programs and advanced technology are also discussed. 1990 (0 471-82584-0) 564 pp. Unified Separation Science J. Calvin Giddings This advanced text/monograph brings together for the first time the variety of techniques used for chemical separations by outlining their common underlying mechanisms. The mass transport phenomena underlying all separation processes are developed in a simple physical-mathematical form, facilitating analysis of alternative separation techniques and the factors integral to separation power. The first six chapters provide background material applicable to a wide range of separation methods, while the final five chapters illustrate specific techniques and methods. 1991 (0 471-52089-6) 320 pp.

Download or read book Size Exclusion Chromatography written by Sadao Mori and published by Springer Science & Business Media. This book was released on 2013-03-09 with total page 236 pages. Available in PDF, EPUB and Kindle. Book excerpt: Although size exclusion chromatography (SEC) is perhaps the most popular and widely used technique for determining the molecular weight distribution of polymeric materials, there have been very few texts written on this topic. During the past decade, SEC has experienced a considerable amount of growth in regard to column and detector technology and new applications. With these advances, SEC can now be used for determining absolute molecular weight, polymer chain conformation and size, and branching, as well as polymer solution properties. This book introduces the reader to the fundamentals of SEC with emphasis on practical aspects of the technique, such as column and mobile selection, calibration, new detector capabilities and guidelines for performing SEC on most types of polymers, especially those of industrial importance. This book is intended for either those new to the field of SEC, or for those research workers who require a more comprehensive background.

Download or read book The Protein Protocols Handbook written by John M. Walker and published by Springer. This book was released on 2007-10-02 with total page 1099 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Protein Protocols Handbook, Second Edition aims to provide a cross-section of analytical techniques commonly used for proteins and peptides, thus providing a benchtop manual and guide for those who are new to the protein chemistry laboratory and for those more established workers who wish to use a technique for the first time. All chapters are written in the same format as that used in the Methods in Molecular BiologyTM series. Each chapter opens with a description of the basic theory behind the method being described. The Materials section lists all the chemicals, reagents, buffers, and other materials necessary for carrying out the protocol. Since the principal goal of the book is to provide experimentalists with a full account of the practical steps necessary for carrying out each protocol successfully, the Methods section contains detailed st- by-step descriptions of every protocol that should result in the successful execution of each method. The Notes section complements the Methods material by indicating how best to deal with any problem or difficulty that may arise when using a given technique, and how to go about making the widest variety of modifications or alterations to the protocol. Since the first edition of this book was published in 1996 there have, of course, been significant developments in the field of protein chemistry.