Choosing the Method of Crystallization to Obtain Optimal Results
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Protocols and Tips in Protein Purification
Department of Molecular Biology & Biotechnology Protocols and tips in protein purification or How to purify protein in one day Second edition 2018 2 Contents I. Introduction 7 II. General sequence of protein purification procedures 9 Preparation of equipment and reagents 9 Preparation and use of stock solutions 10 Chromatography system 11 Preparation of chromatographic columns 13 Preparation of crude extract (cell free extract or soluble proteins fraction) 17 Pre chromatographic steps 18 Chromatographic steps 18 Sequence of operations during IEC and HIC 18 Ion exchange chromatography (IEC) 19 Hydrophobic interaction chromatography (HIC) 21 Gel filtration (SEC) 22 Affinity chromatography 24 Purification of His-tagged proteins 25 Purification of GST-tagged proteins 26 Purification of MBP-tagged proteins 26 Low affinity chromatography 26 III. “Common sense” strategy in protein purification 27 General principles and tips in “common sense” strategy 27 Algorithm for development of purification protocol for soluble over expressed protein 29 Brief scheme of purification of soluble protein 36 Timing for refined purification protocol of soluble over -expressed protein 37 DNA-binding proteins 38 IV. Protocols 41 1. Preparation of the stock solutions 41 2. Quick and effective cell disruption and preparation of the cell free extract 42 3. Protamin sulphate (PS) treatment 43 4. Analytical ammonium sulphate cut (AM cut) 43 5. Preparative ammonium sulphate cut 43 6. Precipitation of proteins by ammonium sulphate 44 7. Recovery of protein from the ammonium sulphate precipitate 44 8. Analysis of solubility of expression 45 9. Analysis of expression for low expressed His tagged protein 46 10. Bio-Rad protein assay Sveta’s easy protocol 47 11. -
Circle Reverse Osmosis System
CIRCLE REVERSE OSMOSIS SYSTEM KEY FEATURES Water Saving Technology – Patented technology eliminates backpressure The RC100 conforms to common in conventional RO systems making the Circle up to 10 times more NSF/ANSI 42, 53 and efficient than existing products. 58 for the reduction of Saves You Money – Conventional RO systems waste up to 24 gallons of Aesthetic Chlorine, Taste water per every 1 gallon of filtered water produced. The Circle only wastes and Odor, Cyst, VOCs, an average of 2.1 gallons of water per 1 gallon of filtered water produced, Fluoride, Pentavalent Arsenic, Barium, Radium 226/228, Cadmium, Hexavalent saving you water and money over the life of the product!. Chromium, Trivalent Chromium, Lead, RO Filter Auto Flushing – Significantly extends life of RO filter. Copper, Selenium and TDS as verified Chrome Faucet Included – With integrated LED filter change indicator. and substantiated by test data. The RC100 conforms to NSF/ANSI 372 for Space Saving Compact Design – Integrated rapid refill tank means more low lead compliance. space under the sink. SPECIFICATIONS Product Name H2O+ Circle Reverse Osmosis Water Filtration System Model / SKU RC100 Installation Undercounter Sediment Filter, Pre-Carbon Plus Filter, Post Carbon Block Filter (RF-20): 6 months Filters & Lifespan RO Membrane Filter (RF-40): 24 months Tank Capacity 6 Liters (refills fully in less than one hour) Dimensions 9.25” (W) x 16.5” (H) x 13.75” (D) Net weight 14.6 lbs Min/Max Operating Pressure 40 psi – 120 psi (275Kpa – 827Kpa) Min/Max Water Feed Temp 41º F – 95º F (5º C – 35º C) Faucet Flow Rate 0.26 – 0.37 gallons per minute (gpm) at incoming water pressure of 20–100 psi Rated Service Flow 0.07 gallons per minute (gpm) Warranty One Year Warranty PO Box 470085, San Francisco CA, 94147–0085 brondell.com 888-542-3355. -
Sedimentation and Clarification Sedimentation Is the Next Step in Conventional Filtration Plants
Sedimentation and Clarification Sedimentation is the next step in conventional filtration plants. (Direct filtration plants omit this step.) The purpose of sedimentation is to enhance the filtration process by removing particulates. Sedimentation is the process by which suspended particles are removed from the water by means of gravity or separation. In the sedimentation process, the water passes through a relatively quiet and still basin. In these conditions, the floc particles settle to the bottom of the basin, while “clear” water passes out of the basin over an effluent baffle or weir. Figure 7-5 illustrates a typical rectangular sedimentation basin. The solids collect on the basin bottom and are removed by a mechanical “sludge collection” device. As shown in Figure 7-6, the sludge collection device scrapes the solids (sludge) to a collection point within the basin from which it is pumped to disposal or to a sludge treatment process. Sedimentation involves one or more basins, called “clarifiers.” Clarifiers are relatively large open tanks that are either circular or rectangular in shape. In properly designed clarifiers, the velocity of the water is reduced so that gravity is the predominant force acting on the water/solids suspension. The key factor in this process is speed. The rate at which a floc particle drops out of the water has to be faster than the rate at which the water flows from the tank’s inlet or slow mix end to its outlet or filtration end. The difference in specific gravity between the water and the particles causes the particles to settle to the bottom of the basin. -
Bottom-Up Self-Assembly Based on DNA Nanotechnology
nanomaterials Review Bottom-Up Self-Assembly Based on DNA Nanotechnology 1, 1, 1 1 1,2,3, Xuehui Yan y, Shujing Huang y, Yong Wang , Yuanyuan Tang and Ye Tian * 1 College of Engineering and Applied Sciences, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210023, China; [email protected] (X.Y.); [email protected] (S.H.); [email protected] (Y.W.); [email protected] (Y.T.) 2 Shenzhen Research Institute of Nanjing University, Shenzhen 518000, China 3 Chemistry and Biomedicine Innovation Center, Nanjing University, Nanjing 210023, China * Correspondence: [email protected] These authors contributed equally to this work. y Received: 9 September 2020; Accepted: 12 October 2020; Published: 16 October 2020 Abstract: Manipulating materials at the atomic scale is one of the goals of the development of chemistry and materials science, as it provides the possibility to customize material properties; however, it still remains a huge challenge. Using DNA self-assembly, materials can be controlled at the nano scale to achieve atomic- or nano-scaled fabrication. The programmability and addressability of DNA molecules can be applied to realize the self-assembly of materials from the bottom-up, which is called DNA nanotechnology. DNA nanotechnology does not focus on the biological functions of DNA molecules, but combines them into motifs, and then assembles these motifs to form ordered two-dimensional (2D) or three-dimensional (3D) lattices. These lattices can serve as general templates to regulate the assembly of guest materials. In this review, we introduce three typical DNA self-assembly strategies in this field and highlight the significant progress of each. -
Optimal Operation of an Integrated Bioreaction–Crystallization Process
中国科技论文在线 http://www.paper.edu.cn Chemical Engineering Science 56 (2001) 6165–6170 www.elsevier.com/locate/ces Optimal operation ofan integrated bioreaction–crystallization process for continuous production of calcium gluconate using external loop airlift columns Jie Baoa, Kenichi Koumatsua, Keiji Furumotob, Makoto Yoshimotoa, Kimitoshi Fukunagaa, Katsumi Nakaoa; ∗ aDepartment of Applied Chemistry and Chemical Engineering, Yamaguchi University, Tokiwadai, Ube, Yamaguchi 755-8611, Japan bOshima National College of Maritime Technology, Oshima, Yamaguchi 742-2106, Japan Abstract A kinetic model was proposed to optimize the integrated bioreaction–crystallization process newly developed for production of calcium gluconate crystals using external loop airlift columns. The optimal operating conditions in the bioreactor were determined using an objective function deÿned to maximize the productivity as well as to minimize biocatalyst loss. The optimization of the crystallizer was carried out by matching the crystallization rate to the optimal production rate in the bioreactor because the bioreaction was found to be the rate controlling process. The calcium gluconate productivity under the optimal conditions of the integrated process was obtained by the simulation based on the process model. The productivity ofthe proposed process was found to be comparable to that ofthe current batch fermentationprocess. ? 2001 Elsevier Science Ltd. All rights reserved. Keywords: Process kinetic model; Optimization; Calcium gluconate; Bioreaction–crystallization -
Anatomy and Physiology of Peritoneal Dialysis
Anatomy and Physiology of Peritoneal Dialysis Isaac Teitelbaum, MD Professor of Medicine Director, Acute & Home Dialysis Programs University of Colorado Hospital Denver, Colorado •1 Outline • Peritoneal cavity as a dialysis system • Models of peritoneal transport • Physiology of peritoneal transport Inverse relationship between solute transport and ultrafiltration • Kinetics of peritoneal transport • Synthesis & Application • Middle Molecules Anatomy of The Peritoneum • The lining of the abdominal cavity • Two layers: parietal - lines the anterior wall and undersurface of the diaphragm - 20% of total SA; blood supply from abdominal wall visceral - covers the abdominal organs - 80% of total SA; blood supply from mesenteric aa and portal vv Gokal R, Textbook of PD, pp. 61-70 •3 Anatomy of The Peritoneum • Size 1.5 – 2 m2; approximates BSA • Highly Vascular • Semi-permeable/bi-directional • “Lymphatic” drainage through diaphragmatic stomata • Continuous with Fallopian Tubes in females Gokal R, Textbook of PD, pp. 61-70 1. The two main properties of the peritoneal membrane are: a. Semi permeable – this allows substances of certain sizes to move from an area of greater concentration to less concentration. b. Bi Directional - substances move in either direction across the membrane. 2. So-called “lymphatic” drainage refers to bulk flow from the peritoneal cavity back to the circulation. This actually occurs across tissues as well as lymphatics. As this is convective flow, dissolved solutes move with the fluid. Thus, fluid reabsorption results in loss of solute clearance as well as loss of fluid removal. 3. It is important to be aware of the continuity of the peritoneal cavity with the Fallopian tubes as retrograde menstruation- which may occur in any woman but goes undetected- will cause bloody dialysate and create concern in the PD patient. -
A Lexicon of Alchemy
A Lexicon of Alchemy by Martin Rulandus the Elder Translated by Arthur E. Waite John M. Watkins London 1893 / 1964 (250 Copies) A Lexicon of Alchemy or Alchemical Dictionary Containing a full and plain explanation of all obscure words, Hermetic subjects, and arcane phrases of Paracelsus. by Martin Rulandus Philosopher, Doctor, and Private Physician to the August Person of the Emperor. [With the Privilege of His majesty the Emperor for the space of ten years] By the care and expense of Zachariah Palthenus, Bookseller, in the Free Republic of Frankfurt. 1612 PREFACE To the Most Reverend and Most Serene Prince and Lord, The Lord Henry JULIUS, Bishop of Halberstadt, Duke of Brunswick, and Burgrave of Luna; His Lordship’s mos devout and humble servant wishes Health and Peace. In the deep considerations of the Hermetic and Paracelsian writings, that has well-nigh come to pass which of old overtook the Sons of Shem at the building of the Tower of Babel. For these, carried away by vainglory, with audacious foolhardiness to rear up a vast pile into heaven, so to secure unto themselves an immortal name, but, disordered by a confusion and multiplicity of barbarous tongues, were ingloriously forced. In like manner, the searchers of Hermetic works, deterred by the obscurity of the terms which are met with in so many places, and by the difficulty of interpreting the hieroglyphs, hold the most noble art in contempt; while others, desiring to penetrate by main force into the mysteries of the terms and subjects, endeavour to tear away the concealed truth from the folds of its coverings, but bestow all their trouble in vain, and have only the reward of the children of Shem for their incredible pain and labour. -
Crystallization in Patterns: a Bio-Inspired Approach**
PROGRESS REPORTS Crystallization in Patterns: A Bio-Inspired Approach** By Joanna Aizenberg* Nature produces a wide variety of exquisite, highly functional mineralized tissues using simple inorganic salts. Biomineralization occurs within specific microenvironments, and is finely tuned by cells and specialized biomacromolecules. This article surveys bio- inspired approaches to artificial crystallization based on the above concept: that is, the use of organized organic surfaces patterned with specific initiation domains on a sub- micrometer scale to control patterned crystal growth. Specially tailored self-assembled monolayers (SAMs) of x-terminated alkanethiols were micropatterned on metal films using soft lithography and applied as organic templates for the nucleation of calcium carbonate. Crystallization results in the formation of large-area, high-resolution inorganic replicas of the underlying organic patterns. SAMs provide sites for ordered nucleation, and make it possible to control various aspects of the crystallization process, including the precise localization of particles, nucleation density, crystal sizes, crystallographic orientation, morphology, polymorph, stability, and architecture. The ability to construct periodic arrays of uniform oriented single crystals, large single crystals with controlled microporosity, or films presenting patterns of crystals offers a potent methodology to materials engineering. 1. Technological Challenge and Biological Of the many challenges facing materials science, the devel- Inspiration opment of an alternative, bottom±up crystallization route, which would enable the direct, patterned growth of crystals The ability to control crystallization is a critical requirement with controlled physico-chemical properties, became an at- in the synthesis of many technologically important materi- tractive, strategic goal. The fundamental principles of the [12±17] als.[1±5] Crystalline inorganic structures with micrometer-scale bottom±up approach can be borrowed from nature. -
CIVL 1101 Introduction to Filtration 1/15
CIVL 1101 Introduction to Filtration 1/15 Water Treatment Water Treatment Water treatment describes those industrial-scale The goal of all water treatment process is to processes used to make water more acceptable remove existing contaminants in the water, or for a desired end-use. reduce the concentration of such contaminants so These can include use for drinking water, industry, the water becomes fit for its desired end-use. medical and many other uses. One such use is returning water that has been used back into the natural environment without adverse ecological impact. Water Treatment Water Treatment Basis water treatment consists Coagulation of four processes: This process helps removes Coagulation/Flocculation particles suspended in water. Sedimentation Chemicals are added to water to form tiny sticky particles Filtration called "floc" which attract the Disinfection particles. Water Treatment Water Treatment Flocculation Sedimentation Flocculation refers to water The heavy particles (floc) treatment processes that settle to the bottom and the combine or coagulate small clear water moves to filtration. particles into larger particles, which settle out of the water as sediment. CIVL 1101 Introduction to Filtration 2/15 Water Treatment Water Treatment Filtration Disinfection The water passes through A small amount of chlorine is filters, some made of layers of added or some other sand, gravel, and charcoal disinfection method is used to that help remove even smaller kill any bacteria or particles. microorganisms that may be in the water. Water Treatment Water Treatment 1. Coagulation 1. Coagulation - Aluminum or iron salts plus chemicals 2. Flocculation called polymers are mixed with the water to make the 3. -
Unsolved Problems in Nanotechnology
Unsolved Problems in Nanotechnology 61 Biographical sketch of Matthew Tirrell Matthew Tirrell received his undergraduate education in Chemical Engineering at Northwestern University and his Ph.D. in 1977 in Polymer Science from the University of Massachusetts. He is currently Dean of the College of Engineering at the University of California, Santa Barbara. From 1977 to 1999 he was on the faculty of Chemical Engineering and Materials Science at the University of Minnesota, where he served as head of the department from 1995 to 1999. His research has been in polymer surface properties including adsorption, adhesion, surface treatment, friction, lubrication and biocompatibilty. He has co-authored about 250 papers and one book and has supervised about 60 Ph.D. students. Professor Tirrell has been a Sloan and a Guggenheim Fellow, a recipient of the Camille and Henry Dreyfus Teacher-Scholar Award and has received the Allan P. Colburn, Charles Stine and the Professional Progress Awards from AIChE. He was elected to the National Academy of Engineering in 1997, became a Fellow of the American Institute of Medical and Biological Engineers in 1998, was elected Fellow of the American Association for the Advancement of Science in 2000 and was named Institute Lecturer for the American Institute of Chemical Engineers in 2001. 62 Unsolved Problems in Nanotechnology: Chemical Processing by Self-Assembly Matthew Tirrell Departments of Chemical Engineering and Materials Materials Research Laboratory California NanoSystems Institute University of California, Santa Barbara, CA 93106-5130 [email protected] Abstract The many impressive laboratory demonstrations of controllable self-assembly methods generate considerable hope and interest in self-assembly as a manufacturing method for nano-structured products. -
The Bio Revolution: Innovations Transforming and Our Societies, Economies, Lives
The Bio Revolution: Innovations transforming economies, societies, and our lives economies, societies, our and transforming Innovations Revolution: Bio The The Bio Revolution Innovations transforming economies, societies, and our lives May 2020 McKinsey Global Institute Since its founding in 1990, the McKinsey Global Institute (MGI) has sought to develop a deeper understanding of the evolving global economy. As the business and economics research arm of McKinsey & Company, MGI aims to help leaders in the commercial, public, and social sectors understand trends and forces shaping the global economy. MGI research combines the disciplines of economics and management, employing the analytical tools of economics with the insights of business leaders. Our “micro-to-macro” methodology examines microeconomic industry trends to better understand the broad macroeconomic forces affecting business strategy and public policy. MGI’s in-depth reports have covered more than 20 countries and 30 industries. Current research focuses on six themes: productivity and growth, natural resources, labor markets, the evolution of global financial markets, the economic impact of technology and innovation, and urbanization. Recent reports have assessed the digital economy, the impact of AI and automation on employment, physical climate risk, income inequal ity, the productivity puzzle, the economic benefits of tackling gender inequality, a new era of global competition, Chinese innovation, and digital and financial globalization. MGI is led by three McKinsey & Company senior partners: co-chairs James Manyika and Sven Smit, and director Jonathan Woetzel. Michael Chui, Susan Lund, Anu Madgavkar, Jan Mischke, Sree Ramaswamy, Jaana Remes, Jeongmin Seong, and Tilman Tacke are MGI partners, and Mekala Krishnan is an MGI senior fellow. -
Membrane Filtration
Membrane Filtration A membrane is a thin layer of semi-permeable material that separates substances when a driving force is applied across the membrane. Membrane processes are increasingly used for removal of bacteria, microorganisms, particulates, and natural organic material, which can impart color, tastes, and odors to water and react with disinfectants to form disinfection byproducts. As advancements are made in membrane production and module design, capital and operating costs continue to decline. The membrane processes discussed here are microfiltration (MF), ultrafiltration (UF), nanofiltration (NF), and reverse osmosis (RO). MICROFILTRATION Microfiltration is loosely defined as a membrane separation process using membranes with a pore size of approximately 0.03 to 10 micronas (1 micron = 0.0001 millimeter), a molecular weight cut-off (MWCO) of greater than 1000,000 daltons and a relatively low feed water operating pressure of approximately 100 to 400 kPa (15 to 60psi) Materials removed by MF include sand, silt, clays, Giardia lamblia and Crypotosporidium cysts, algae, and some bacterial species. MF is not an absolute barrier to viruses. However, when used in combination with disinfection, MF appears to control these microorganisms in water. There is a growing emphasis on limiting the concentrations and number of chemicals that are applied during water treatment. By physically removing the pathogens, membrane filtration can significantly reduce chemical addition, such as chlorination. Another application for the technology is for removal of natural synthetic organic matter to reduce fouling potential. In its normal operation, MF removes little or no organic matter; however, when pretreatment is applied, increased removal of organic material can occur.