DOCSLIB.ORG

Controlling Nonspecific Adsorption of Proteins at Bio-Interfaces for Biosensor and Biomedical Applications

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Controlling Nonspecific Adsorption of Proteins at Bio-Interfaces for Biosensor and Biomedical Applications Utah State University DigitalCommons@USU All Graduate Theses and Dissertations Graduate Studies 5-2009 Controlling Nonspecific Adsorption of Proteins at Bio-Interfaces for Biosensor and Biomedical Applications Harshil D. Dhruv Utah State University Follow this and additional works at: https://digitalcommons.usu.edu/etd Part of the Biomedical Engineering and Bioengineering Commons Recommended Citation Dhruv, Harshil D., "Controlling Nonspecific Adsorption of Proteins at Bio-Interfaces for Biosensor and Biomedical Applications" (2009). All Graduate Theses and Dissertations. 276. https://digitalcommons.usu.edu/etd/276 This Dissertation is brought to you for free and open access by the Graduate Studies at DigitalCommons@USU. It has been accepted for inclusion in All Graduate Theses and Dissertations by an authorized administrator of DigitalCommons@USU. For more information, please contact [email protected]. CONTROLLING NONSPECIFIC ADSORPTION OF PROTEINS AT BIO- INTERFACES FOR BIOSENSOR AND BIOMEDICAL APPLICATIONS by Harshil D. Dhruv A dissertation submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in Biological Engineering Approved: __________________________ __________________________ David W. Britt Anhong Zhou Committee Chairman Committee Member __________________________ __________________________ Soonjo Kwon Marie K. Walsh Committee Member Committee Member __________________________ __________________________ Silvana Martini Byron R. Burnham Committee Member Dean of Graduate Studies UTAH STATE UNVERSITY Logan, Utah 2009 ii Copyright © Harshil D. Dhruv 2009 All Rights Reserved iii ABSTRACT Controlling Nonspecific Adsorption of Proteins at Bio-interfaces for Biosensor and Biomedical Applications by Harshil D. Dhruv, Doctor of Philosophy Utah State University, 2009 Major Professor: Dr. David W. Britt Department: Biological and Irrigation Engineering Partitioning of poly(ethyleneglycol) (PEG) molecules in 2-D and 3-D systems is presented as a self-assembly approach for controlling non-specific adsorption of proteins at interfaces. Lateral restructuring of multi-component Langmuir monolayers to accommodate adsorbing proteins was investigated as a model 2-D system. Ferritin adsorption to monolayers containing cationic, nonionic, and PEG bearing phospholipids induced protein sized binding pockets surrounded by PEG rich regions. The number, size, and distribution of protein imprint sites were controlled by the molar ratios, miscibility, and lateral mobility of the lipids. The influence of PEG chain length on the ternary monolayer restructuring and protein distribution was also investigated using DSPE-PEGx (x= 7, 16, 22). Monolayer miscibility analysis demonstrated that longer PEG chains diminished the condensed phase formation for a fixed ratio of lipids. Thus, incorporation of longer PEG chains, iv intended to diminish protein adsorption outside of the imprint sites of cationic / non-ionic lipids, leads to dramatic changes in monolayer phase behavior and protein distribution in this 2-D system. The assembly of PEG-amphiphiles at elastomer surfaces and subsequent protein adsorption was investigated as a model 3-D system. Polydimethylsiloxane (PDMS) substrates were modified with block copolymers comprised of PEG and PDMS segments by two methods: (1) the block copolymer was mixed with PDMS during polymerization; (2) the block copolymer diffused into solvent swollen PDMS monoliths. Hydrophilic surfaces resulted for both approaches that, for 600 D block copolymer, exhibited up to 85% reduction in fibrinogen adsorption as compared to native PDMS. Higher MW block copolymers (up to 3000 D) resulted in less hydrophilic surfaces and greater protein adsorption, presumably due to diffusion limitations of copolymer in the PDMS monolith. All modified PDMS surfaces were dynamic and restructured when cycled between air and water. PDMS transparency also decreased with increase in block copolymer concentration for both methods, limiting this modification protocol for applications requiring high polymer transparency. The 2-D system presents a bottom-up approach, where adsorbing protein constructs the binding site, while the 3-D system presents a top down approach, where protein-binding elements may be introduced into the PEG-bearing polymer for fabrication of surfaces with controlled protein adsorption. (220 pages) v To my parents Dinesh B. Dhruv and Poornima D. Dhruv and my loving wife Nikita Zaveri vi ACKNOWLEDGMENTS I would like to express my deepest gratitude to my professor and mentor, Dr. David W. Britt, for giving me the opportunity to dream, create, and discover in his laboratory during my PhD work. I would also like to thank other professors on my committee, including Dr. Marie K. Walsh, Dr. Anhong Zhou, Dr. Silvana Martini, and Dr. Soonjo Kwon, for their valuable support and advice during the course of my work. I am heartily thankful to the late Dr. Lyman S. Willardson for giving me the opportunity to study at Utah State University. I am thankful to all my colleagues at Dr. Britt’s lab for their help and discussion in improving my dissertation. I gratefully acknowledge the help given by Mr. Bradley Tuft for collecting stability study data for PDMS samples. I am also thankful to all my friends at Utah State University for giving me a friendly environment during the course of my studies. Finally, I would like to thank my parents, my sister, and my wife for their continuous support and encouragement they have provided me during these years, without which this journey would not have been possible for me. Harshil Dhruv vii CONTENTS Page ABSTRACT....................................................................................................................... iii DEDICATION................................................................................................................... vi ACKNOWLEDGMENTS ................................................................................................ vii LIST OF TABLES............................................................................................................ xii LIST OF FIGURES ......................................................................................................... xiii LIST OF SCHEMES....................................................................................................... xxii LIST OF SYMBOLS, NOTATIONS, AND DEFINITIONS........................................ xxiii CHAPTER 1 INTRODUCTION ...................................................................................................1 1.1 Background...............................................................................................1 1.2 Hypothesis and Objectives........................................................................3 1.3 Dissertation Outline..................................................................................4 1.4 References.................................................................................................5 2 LITERATURE REVIEW........................................................................................8 2.1 Protein Adsorption to Biologically Relevant Surfaces .............................8 2.1.1 Specific Protein Adsorption..........................................................9 2.1.2 Non-specific Protein Adsorption ................................................10 2.1.2.1 Causes of Non-specific Protein Adsorption.................10 2.1.2.2 Consequences of Non-specific Protein Adsorption....................................................................14 2.2 Surface Modification to Prevent Non-specific Protein Adsorption........17 2.3 Protein Repellent/Nonfouling Coating Materials ...................................18 2.3.1 Biomemetic Materials.................................................................19 2.3.1.1 Phosphorylcholines (PC).............................................20 viii 2.3.1.2 Oligo/Polysaccharides..................................................21 2.3.2 Synthetic Polymers .....................................................................23 2.3.2.1 Polyacrylates................................................................23 2.3.2.2 Oligo- and Poly(ethyleneglycol)s ................................25 2.4 Poly(ethyleneglycol) (PEG)....................................................................25 2.4.1 PEG Properties............................................................................26 2.4.1.1 The Physical View of Fouling Resistance ...................27 2.4.1.2 The Chemical View of Fouling Resistance .................28 2.5 Traditional PEG Surface Immobilization Methods ................................30 2.5.1 Graft Copolymerization ..............................................................31 2.5.2 Chemical Coupling.....................................................................35 2.5.3 Non-covalent Immobilization .....................................................38 2.5.3.1 Physisorption................................................................39 2.5.3.2 Langmuir and Langmuir-Blodgett (LB) Technique.....................................................................40 2.5.3.2.1 Langmuir Monolayers................................40 2.5.3.2.2 Langmuir-Blodgett (LB) Films..................43 2.5.3.2.3 Langmuir and LB Films for Protein Repellent Coatings.....................................44 2.6 Langmuir and LB Films for Protein Patterning ......................................45 2.7 Poly(Dimethylsiloxane) (PDMS)............................................................49
Load more

Recommended publications
  • MEMS Technology for Physiologically Integrated Devices
    A BioMEMS Review: MEMS Technology for Physiologically Integrated Devices AMY C. RICHARDS GRAYSON, REBECCA S. SHAWGO, AUDREY M. JOHNSON, NOLAN T. FLYNN, YAWEN LI, MICHAEL J. CIMA, AND ROBERT LANGER Invited Paper MEMS devices are manufactured using similar microfabrica- I. INTRODUCTION tion techniques as those used to create integrated circuits. They often, however, have moving components that allow physical Microelectromechanical systems (MEMS) devices are or analytical functions to be performed by the device. Although manufactured using similar microfabrication techniques as MEMS can be aseptically fabricated and hermetically sealed, those used to create integrated circuits. They often have biocompatibility of the component materials is a key issue for moving components that allow a physical or analytical MEMS used in vivo. Interest in MEMS for biological applications function to be performed by the device in addition to (BioMEMS) is growing rapidly, with opportunities in areas such as biosensors, pacemakers, immunoisolation capsules, and drug their electrical functions. Microfabrication of silicon-based delivery. The key to many of these applications lies in the lever- structures is usually achieved by repeating sequences of aging of features unique to MEMS (for example, analyte sensitivity, photolithography, etching, and deposition steps in order to electrical responsiveness, temporal control, and feature sizes produce the desired configuration of features, such as traces similar to cells and organelles) for maximum impact. In this paper, (thin metal wires), vias (interlayer connections), reservoirs, we focus on how the biological integration of MEMS and other valves, or membranes, in a layer-by-layer fashion. The implantable devices can be improved through the application of microfabrication technology and concepts.
    [Show full text]
  • Cell and Protein Compatibility of Parylene-C Surfaces
    Cell and Protein Compatibility of Parylene-C Surfaces Tracy Y. Chang,†,# Vikramaditya G. Yadav,‡,¶,# Sarah De Leo,§ Agustin Mohedas,⊥ Bimal Rajalingam,£ Chia-Ling Chen,| Selvapraba Selvarasah,| Mehmet R. Dokmeci,| and Ali Khademhosseini*,£,¶ Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, Department of Chemical Engineering, UniVersity of Waterloo, Waterloo, Canada N2L 3G1, Department of Biological Engineering, Louisiana State UniVersity, Baton Rouge, Louisiana 70808, Department of Biomedical Engineering, Texas A&M UniVersity, College Station, Texas 77843, Center for Biomedical Engineering, Department of Medicine, Brigham and Women’s Hospital, HarVard Medical School, Boston, Massachusetts 02139, Department of Electrical and Computer Engineering, Northeastern UniVersity, Boston, Massachusetts 02115, and HarVard-MIT DiVision of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 ReceiVed June 11, 2007. In Final Form: August 9, 2007 Parylene-C, which is traditionally used to coat implantable devices, has emerged as a promising material to generate miniaturized devices due to its unique mechanical properties and inertness. In this paper we compared the surface properties and cell and protein compatibility of parylene-C relative to other commonly used BioMEMS materials. We evaluated the surface hydrophobicity and roughness of parylene-C and compared these results to those of tissue culture-treated polystyrene, poly(dimethylsiloxane) (PDMS), and glass. We also treated parylene-C and PDMS with air plasma, and coated the surfaces with fibronectin to demonstrate that biochemical treatments modify the surface properties of parylene-C. Although plasma treatment caused both parylene-C and PDMS to become hydrophilic, only parylene-C substrates retained their hydrophilic properties over time. Furthermore, parylene-C substrates display a higher degree of nanoscale surface roughness (>20 nm) than the other substrates.
    [Show full text]
  • Proteins and Peptides As Important Modifiers of the Polymer Scaffolds
    polymers Review Proteins and Peptides as Important Modifiers of the Polymer Scaffolds for Tissue Engineering Applications—A Review Katarzyna Klimek * and Grazyna Ginalska Chair and Department of Biochemistry and Biotechnology, Medical University of Lublin, Chodzki 1 Street, 20-093 Lublin, Poland; [email protected] * Correspondence: [email protected]; Tel.: +48-81-448-7028; +48-81-448-7020 Received: 29 January 2020; Accepted: 2 April 2020; Published: 6 April 2020 Abstract: Polymer scaffolds constitute a very interesting strategy for tissue engineering. Even though they are generally non-toxic, in some cases, they may not provide suitable support for cell adhesion, proliferation, and differentiation, which decelerates tissue regeneration. To improve biological properties, scaffolds are frequently enriched with bioactive molecules, inter alia extracellular matrix proteins, adhesive peptides, growth factors, hormones, and cytokines. Although there are many papers describing synthesis and properties of polymer scaffolds enriched with proteins or peptides, few reviews comprehensively summarize these bioactive molecules. Thus, this review presents the current knowledge about the most important proteins and peptides used for modification of polymer scaffolds for tissue engineering. This paper also describes the influence of addition of proteins and peptides on physicochemical, mechanical, and biological properties of polymer scaffolds. Moreover, this article sums up the major applications of some biodegradable natural and synthetic polymer scaffolds modified with proteins and peptides, which have been developed within the past five years. Keywords: bioactive construct; biocompatibility; biomolecules; cytotoxicity; ECM; hydrogels; protein carrier; regenerative medicine; stem cells; tissue repair 1. Introduction: The Role of Proteins and Peptides in TE Tissue engineering (TE) is a multidisciplinary field, which constitutes an alternative and promising approach for grafts, i.e., autografts, allografts, and xenografts [1–3].
    [Show full text]
  • Characterization of the Adsorption of Biomolecules in Open Electrowetting on Dielectrics Microfluidics-Tool Platform for Bio-Analytical Applications
    Characterization of the Adsorption of Biomolecules in Open Electrowetting On Dielectrics Microfluidics-Tool Platform for Bio-analytical Applications OEWOD surfaces for biomolecular adsorption experiments i Characterization of the Adsorption of Biomolecules in Open Electrowetting On Dielectrics Microfluidics-Tool Platform for Bio-analytical Applications Miguel Angel Martínez Garza Born in Mexico City A thesis presented in fulfilment of the requirement for the degree of Doctor of philosophy Institute of Microsystems Technology Department of Sensors Albert-Ludwig University Freiburg, Germany August 2011 ii Dekan: Prof. Dr. Bernd Becker Prüfungskommission: Vorsitz: Prof. Rühe Beisitz: Prof. Stieglitz Gutachter: Prof. Urban und Prof. Woias Datum der Prüfung: 02.03.12 iii Summary Open Electrowetting On Dielectrics (OEWOD) is one of several designs of the original Electrowetting On Dielectrics (EWOD) microfluidics platform. EWOD has gained considerable attention for its capacity of transporting smallest volumes of liquids especially in biochips and BioMEMS approaches. The electrical polarization of the dielectrics layer causing the apparent increase in surface energy is uses to transport microdroplets to carry out the basic operations fulfilling a bioassay protocol. This thesis was mainly focused to describe and characterize the behaviour of several electrolytes and biomolecules solution under OEWOD conditions. A general overview of OEWOD microfluidics platform will provide by (1) the characterization by Contact Angle (CA). (2) the atomic Force Microscopy (AFM) of OEWOD surfaces. (3) the Laser Scanning Confocal Microscopy (LSCM) of OEWOD surfaces, and (4) the relative quantitative biomolecular adsorption behaviour of several biomolecules as Horseradish Peroxidase [HRP], Peroxidase-Conjugated Rabbit Anti-Mouse Immunoglobulin [HRP-IgG] and Deoxyribonucleic acid-Peroxidase [HRP-DNA] under OEWOD ―actuation‖ and static conditions.
    [Show full text]
  • Titanium and Protein Adsorption: an Overview of Mechanisms and Effects of Surface Features
    materials Review Titanium and Protein Adsorption: An Overview of Mechanisms and Effects of Surface Features Jacopo Barberi * and Silvia Spriano Department of Applied Science and Technology, Politecnico di Torino, 10129 Turin, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-011-0904567 Abstract: Titanium and its alloys, specially Ti6Al4V, are among the most employed materials in orthopedic and dental implants. Cells response and osseointegration of implant devices are strongly dependent on the body–biomaterial interface zone. This interface is mainly defined by proteins: They adsorb immediately after implantation from blood and biological fluids, forming a layer on implant surfaces. Therefore, it is of utmost importance to understand which features of biomaterials surfaces influence formation of the protein layer and how to guide it. In this paper, relevant literature of the last 15 years about protein adsorption on titanium-based materials is reviewed. How the surface characteristics affect protein adsorption is investigated, aiming to provide an as comprehensive a picture as possible of adsorption mechanisms and type of chemical bonding with the surface, as well as of the characterization techniques effectively applied to model and real implant surfaces. Surface free energy, charge, microroughness, and hydroxylation degree have been found to be the main surface parameters to affect the amount of adsorbed proteins. On the other hand, the conformation of adsorbed proteins is mainly dictated by the protein structure, surface topography at the nano-scale, and exposed functional groups. Protein adsorption on titanium surfaces still needs further clarification, in particular concerning adsorption from complex protein solutions. In addition, characterization techniques to investigate and compare the different aspects of protein adsorption on Citation: Barberi, J.; Spriano, S.
    [Show full text]
  • Adsorption of Proteins at Solid-Liquid Interfaces
    Cells and Materials Volume 5 Number 1 Article 9 1995 Adsorption of Proteins at Solid-Liquid Interfaces Willem Norde Wageningen Agricultural University, The Netherlands Follow this and additional works at: https://digitalcommons.usu.edu/cellsandmaterials Part of the Biomedical Engineering and Bioengineering Commons Recommended Citation Norde, Willem (1995) "Adsorption of Proteins at Solid-Liquid Interfaces," Cells and Materials: Vol. 5 : No. 1 , Article 9. Available at: https://digitalcommons.usu.edu/cellsandmaterials/vol5/iss1/9 This Article is brought to you for free and open access by the Western Dairy Center at DigitalCommons@USU. It has been accepted for inclusion in Cells and Materials by an authorized administrator of DigitalCommons@USU. For more information, please contact [email protected]. Cells and Materials, Vol. 5, No. 1, 1995 (Pages 97-112) 1051-6794/95$5.00+ .25 Scanning Microscopy International , Chicago (AMF O'Hare), IL 60666 USA ADSORPTION OF PROTEINS AT SOLID-LIQillD INTERFACES Willem Norde Department of Physical and Colloid Chemi stry, Wageningen Agri cultural Universi ty, P.O. Box 8038, 6700 EK Wageningen, The Netherlands Telephone number: (31 ) 08370-83540 I FAX number: (31 ) 08370-83777 (Received for publicati on October 4, 1994 and in revised form March 31, 1995) Abstract Introduction This paper concisely reviews the general principles Interacti on between proteins and solid surfaces is underl ying protein adsorption from aqueous solution commonly observed, both in natural and syntheti c sys­ onto a solid surface. The discussion includes the various tems. These interactions are of great relevance in , e.g., stages of the adsorption process, i. e., transport of the medical, bi otechnological and envi ronmental applica­ protein molecules towards the surface, the absorbed tions.
    [Show full text]
  • Force Microscopy Studies of Fibronectin Adsorption and Subsequent Cellular Adhesion to Substrates with Well-Defined Surface Chemistries
    Force Microscopy Studies of Fibronectin Adsorption and Subsequent Cellular Adhesion to Substrates With Well-Defined Surface Chemistries Pamela Meadows, Gilbert Walker Abstract Molecular force spectroscopy was used to study the mechanical behavior of plasma fibronectin (FN) on mica, gold, poly(ethylene) glycol (PEG), and -CH3, -OH, and -COOH terminated alkanethiol self-assembled monolayers. Proteins were examined at two concentrations, one resulting in a saturated surface with multiple intermolecular interactions referred to as the aggregate state and another resulting in a semiaggregate state where the proteins were neither completely isolated nor completely aggregated. Modeling of the force-extension data using two different theories resulted in similar trends for the fitted thermodynamic parameters from which insight into the protein’s binding state could be obtained. Aggregated proteins adsorbed on hydrophobic surfaces adopted more rigid conformations apparently as a result of increased surface denaturation and tighter binding while looser conformations were observed on more hydrophilic surfaces. Introduction Fibronectin (FN) is a large multidomain protein that is found on cell surfaces, in plasma, and other body fluids as well as being a major constituent of the extracellular matrix (ECM). Of particular interest is the elasticity of this protein. It is believed to be crucial for the formation of fibronectin fibrils in the ECM which in turn can increase cell and molecular adhesion to the surface. In this study, atomic force microscopy is being used to study fibronectin’s elasticity to obtain new insights into its structural properties and how these properties are influenced by the surface. By understanding how the surface influences the protein’s conformation, new biomaterials can then be designed.
    [Show full text]
  • An Investigation of the Mechanisms of Electronic Sensing of Protein Adsorption on Carbon Nanotube Devices Robert J
    Published on Web 01/15/2004 An Investigation of the Mechanisms of Electronic Sensing of Protein Adsorption on Carbon Nanotube Devices Robert J. Chen,†,§ Hee Cheul Choi,†,§,| Sarunya Bangsaruntip,† Erhan Yenilmez,† Xiaowu Tang,† Qian Wang,† Ying-Lan Chang,‡ and Hongjie Dai*,† Contribution from the Department of Chemistry, Stanford UniVersity, Stanford, California 94305, and Agilent Laboratories, Agilent Technologies, Inc., 3500 Deer Creek Road, Palo Alto, California 94304 Received September 24, 2003; E-mail: [email protected] Abstract: It has been reported that protein adsorption on single-walled carbon nanotube field effect transistors (FETs) leads to appreciable changes in the electrical conductance of the devices, a phenomenon that can be exploited for label-free detection of biomolecules with a high potential for miniaturization. This work presents an elucidation of the electronic biosensing mechanisms with a newly developed microarray of nanotube “micromat” sensors. Chemical functionalization schemes are devised to block selected components of the devices from protein adsorption, self-assembled monolayers (SAMs) of methoxy(poly- (ethylene glycol))thiol (mPEG-SH) on the metal electrodes (Au, Pd) and PEG-containing surfactants on the nanotubes. Extensive characterization reveals that electronic effects occurring at the metal-nanotube contacts due to protein adsorption constitute a more significant contribution to the electronic biosensing signal than adsorption solely along the exposed lengths of the nanotubes. Introduction suitable
    [Show full text]
  • Quantitative Comparison of Protein Adsorption and Conformational Changes on Dielectric‑Coated Nanoplasmonic Sensing Arrays
    This document is downloaded from DR‑NTU (https://dr.ntu.edu.sg) Nanyang Technological University, Singapore. Quantitative comparison of protein adsorption and conformational changes on dielectric‑coated nanoplasmonic sensing arrays Jackman, Joshua A.; Sut, Tun Naw; Cho, Nam‑Joon; Ferhan, Abdul Rahim 2018 Ferhan, A. R., Jackman, J. A., Sut, T. N., & Cho, N.‑J. (2018). Quantitative comparison of protein adsorption and conformational changes on dielectric‑coated nanoplasmonic sensing arrays. Sensors, 18(4), 1283‑. https://hdl.handle.net/10356/87502 https://doi.org/10.3390/s18041283 © 2018 by The Author(s). Licensee MDPI, Basel, Switzerland. This article is an open accessarticle distributed under the terms and conditions of the Creative Commons Attribution(CC BY) license (http://creativecommons.org/licenses/by/4.0/). Downloaded on 24 Sep 2021 23:57:31 SGT sensors Article Quantitative Comparison of Protein Adsorption and Conformational Changes on Dielectric-Coated Nanoplasmonic Sensing Arrays Abdul Rahim Ferhan 1, Joshua A. Jackman 1, Tun Naw Sut 1 and Nam-Joon Cho 1,2,* ID 1 School of Materials Science and Engineering and Centre for Biomimetic Sensor Science, Nanyang Technological University, 50 Nanyang Drive 637553, Singapore; [email protected] (A.R.F.); [email protected] (J.A.J.); [email protected] (T.N.S.) 2 School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive 637459, Singapore * Correspondence: [email protected]; Tel.: +65-6790-4925 Received: 16 March 2018; Accepted: 19 April 2018; Published: 22 April 2018 Abstract: Nanoplasmonic sensors are a popular, surface-sensitive measurement tool to investigate biomacromolecular interactions at solid-liquid interfaces, opening the door to a wide range of applications.
    [Show full text]
  • The Influence of Amino Acid Properties on The
    THE INFLUENCE OF AMINO ACID PROPERTIES ON THE ADSORPTION OF PROTEINS AND PEPTIDES TO STAINLESS STEEL SURFACES A thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy in Chemical and Process Engineering University of Canterbury New Zealand NEHA CHANDRASEKARAN March 2014 To my beloved Master, spiritual guide, teacher and friend Baba Gurinder Singh Dhillon i Abstract Stainless steel (SS) is the material of choice in a number of process industries ranging from food and dairy to pharmaceuticals. Adsorption phenomena on SS surfaces are of paramount importance in these industries. For example, protein adsorption constitutes a major issue in process equipment, as the associated surface fouling decreases the efficiency of the overall process and leads to an increase in operational costs because of the need for regular cleaning. In addition, the adsorption of proteins at solid–liquid interfaces is an important research field with relevance in biosensor and biomaterial applications. The primary aim of this thesis was to understand the underlying adsorption properties of selected protein onto SS surfaces and to identify the influence of specific amino acids on bio-fouling. Protein adsorption experiments were carried out on 316 grade SS sensors using a quartz crystal microbalance with dissipation (QCM-D). The proteins consisted of milk proteins (α-lactalbumin, β-lactoglobulin, α-casein, β-casein, κ-casein and bovine serum albumin), blood proteins (cytochrome-c, haemoglobin and myoglobin) and proteins of industrial and medical relevance (α- chymotrypsinogen, human recombinant insulin, lysozyme and papain). The adsorption characteristics of the test proteins were studied and an empirical correlation relating the amount of protein adsorbed to their physical properties was proposed.
    [Show full text]
  • Studies of Protein Adsorption on Implant Materials in Relation to Biofilm Formation
    Studies of protein adsorption on implant materials in relation to biofilm formation I. Activity of Pseudomonas aeruginosa on Polypropylene and High density Polyethylene in presence of serum albumin. Authors: S Dutta Sinha*1, Susmita Chatterjee3, P.K.Maity3, S.Tarafdar1, S.P.Moulik2 Abstract: The surface of biomaterials used as implants are highly susceptible to bacterial colonization and subsequent infection. The amount of protein adsorption on biomaterials, among other factors, can affect the nature and quality of biofilms formed on them. The variation in the adsorption time of the protein on the biomaterial surface produces a phenotypic change in the bacteria by alteration of the production of EPS (exoplysaccharide) matrix. Knowledge of the effects of protein adsorption on implant infection will be very useful in understanding the chemistry of the biomaterial surfaces, which can deter the formation of biofilms. It is observed that the adsorption of BSA on the biomaterial surfaces increases with time and concentration, irrespective of their type and the nature of the EPS matrix of the bacterial biofilm is dependent on the amount of protein adsorbed on the biomaterial surface. The adsorption of protein (BSA) on the biomaterials, polypropylene (PP) and high density polyethylene (HDPE) has been studied and the formation of the biofilms of Pseudomonas aeruginosa on them has been examined. 1. Introduction Infections associated with implanted devices are notoriously difficult to treat and often lead to serious complications. [1, 2, 3] As has been widely discussed,[4,5,6] the core issue of implant infection involves bacterial attachment and colonization on solid surfaces, which can be otherwise termed as ‘biofilm formation’.
    [Show full text]
  • Characterization of Protein Surface Interactions: Collagen and Osteocalcin
    Department of Physics, Chemistry and Biology Master's Thesis Characterization of Protein Surface Interactions: Collagen and Osteocalcin Patrik Johansson 2013-06-17 LITH-IFM-A-EX--13/2780--SE Linköping University Department of Physics, Chemistry and Biology 581 83 Linköping Department of Physics, Chemistry and Biology Characterization of Protein Surface Interaction: Collagen and Osteocalcin Patrik Johansson Thesis work done at NESAC/BIO, University of Washington, Seattle. 2013-06-17 Supervisor Prof. Patrick Koelsch Examiner Prof. Kajsa Uvdal Linköping University Department of Physics, Chemistry and Biology 581 83 Linköping 2 Avdelning, institution Datum Division, Department Date Physics 2013-06-17 Department of Physics, Chemistry and Biology Linköping University Språk Rapporttyp ISBN Language Report category Svenska/Swedish Licentiatavhandling ISRN: LITH-IFM-A-EX--13/2780--SE Engelska/English Examensarbete _________________________________________________________________ C-uppsats D-uppsats Serietitel och serienummer ISSN ________________ Övrig rapport Title of series, numbering ______________________________ _____________ URL för elektronisk version Titel Title Characterization of Protein Surface Characterization: Collagen and Osteocalcin Författare Author Patrik Johansson Sammanfattning Abstract This work investigates how the proteins collagen type I and human osteocalcin interact with various surfaces. A pH- series of collagen adsorbed onto methyl terminated self-assembled monolayers has been made and the results indicate that less tropocollagen is found on the surfaces at pH below 6.0 and that biofilms made of larger fibrils with a more ordered 3D-structure are formed at pH 6.0 and above. This work also shows that it is possible to divide the amide I region of a vibrational Sum Frequency Generation (v-SFG) spectra into three peaks.
    [Show full text]