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Detection and Thermal Stabilization of Virus Based on Surface Properties Michigan Technological University Digital Commons @ Michigan Tech Dissertations, Master's Theses and Master's Reports 2020 Detection and thermal stabilization of virus based on surface properties Xue Mi Michigan Technological University, [email protected] Copyright 2020 Xue Mi Recommended Citation Mi, Xue, "Detection and thermal stabilization of virus based on surface properties", Open Access Dissertation, Michigan Technological University, 2020. https://doi.org/10.37099/mtu.dc.etdr/1020 Follow this and additional works at: https://digitalcommons.mtu.edu/etdr Part of the Biochemical and Biomolecular Engineering Commons DETECTION AND THERMAL STABILIZATION OF VIRUS BASED ON SURFACE PROPERTIES By Xue Mi A DISSERTATION Submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY In Chemical Engineering MICHIGAN TECHNOLOGICAL UNIVERSITY 2020 © 2020 Xue Mi This dissertation has been approved in partial fulfillment of the requirements for the Degree of DOCTOR OF PHILOSOPHY in Chemical Engineering. Department of Chemical Engineering Dissertation Advisor: Dr. Caryn Heldt Committee Member: Dr. Adrienne Minerick Committee Member: Dr. Lei Pan Committee Member: Dr. Marina Tanasova Department Chair: Dr. Pradeep K. Agrawal Table of Contents List of Figures ................................................................................................................... vii List of Tables .................................................................................................................... xii Preface.............................................................................................................................. xiii Acknowledgements .......................................................................................................... xiv Abstract ..............................................................................................................................xv 1 Introduction and overview ..........................................................................................1 1.1 Introduction ......................................................................................................2 1.2 Overview ..........................................................................................................3 1.3 Reference ..........................................................................................................5 2 Literature review .........................................................................................................7 2.1 General overview of viral diseases and viruses ................................................8 2.1.1 Viral diseases outbreak .......................................................................8 2.1.2 Viral structure and families .................................................................9 2.2 Virus characterization .....................................................................................11 2.2.1 Virus surface properties ....................................................................11 2.2.1.1 Hydrophobicity ...............................................................12 2.2.1.2 Surface charge and isoelectric point (pI) ........................13 2.2.2 Traditional virus surface characterization methods ..........................14 2.2.2.1 Hydrophobic interaction chromatography (HIC) ............14 2.2.2.2 Aqueous two-phase system (ATPS) partitioning method 16 2.2.2.3 Zeta potential ..................................................................17 2.2.2.4 Isoelectric focusing (IEF) ...............................................21 2.2.3 Novel surface characterization: chemical force microscopy (CFM) 22 2.2.3.1 Fundamental principles of atomic force microscope (AFM) 23 2.2.3.2 Force measurement (approach curve vs. withdraw curve) 25 2.3 Virus detection ...............................................................................................30 2.3.1 Past and current virus detection methods ..........................................30 2.3.1.1 Transmission electron microscopy (TEM) .....................30 2.3.1.2 Immunoassays .................................................................31 2.3.1.3 Polymerase chain reaction ..............................................33 2.3.1.4 In vitro cell culture assays...............................................34 2.3.2 Gold nanoparticles (AuNPs) in virus detection ................................36 iii 2.3.2.1 AuNPs properties ............................................................36 2.3.2.2 Detection of specific viruses using AuNPs .....................37 2.4 Virus thermal stability ....................................................................................42 2.4.1 Viral based vaccines .........................................................................42 2.4.2 Cold chain and cold storage problem................................................42 2.4.3 Conventional vaccine formulation to stabilize viral vaccines ..........44 2.4.4 Novel formulations to enhance the thermal stability of virus by complex coacervation .....................................................................................46 2.5 References ......................................................................................................51 3 Virus surface chemistry characterization ..................................................................64 3.1 Introduction ....................................................................................................65 3.2 Experimental section ......................................................................................66 3.2.1 Materials ...........................................................................................66 3.2.2 Virus purification ..............................................................................67 3.2.3 Virus samples and control surfaces preparation ...............................67 3.2.4 AFM probe modification ..................................................................67 3.2.5 AFM imaging, force measurement and analysis ...............................68 3.3 Results and discussion ....................................................................................68 3.3.1 Virus and virus surrogate immobilization.........................................69 3.3.2 AFM probe modification ..................................................................72 3.3.3 Virus surface probing by CFM .........................................................75 3.4 Conclusions ....................................................................................................77 3.5 References ......................................................................................................78 4 Virus isoelectric point determination ........................................................................82 4.1 Introduction ....................................................................................................83 4.2 Experimental section ......................................................................................85 4.2.1 Materials. ..........................................................................................85 4.2.2 Buffers...............................................................................................85 4.2.3 Virus production, purification, and titration. ....................................85 4.2.4 Virus samples and control surfaces preparation. ..............................86 4.2.5 AFM probe functionalization. ...........................................................86 4.2.6 AFM imaging, force measurement, and analysis. .............................87 4.2.7 Zeta potential measurement. .............................................................87 4.2.8 ATPS measurement. .........................................................................87 4.2.9 Theoretical calculation of virus surface potential and pI. .................88 4.3 Results and discussion ....................................................................................88 4.3.1 Surface and probe preparations. ........................................................88 4.3.2 Virus pI using CFM. .........................................................................91 iv 4.3.3 Comparison to theoretically calculated virus pI and surface potential. 94 4.4 Conclusions ....................................................................................................96 4.5 References ......................................................................................................97 5 Virus detection by mannitol-induced gold nanoparticle aggregation .....................101 5.1 Introduction ..................................................................................................102 5.2 Experimental section ....................................................................................104 5.2.1 Materials .........................................................................................104 5.2.2 Virus titration and purification ........................................................104 5.2.3 Synthesis and characterization of AuNPs .......................................104 5.2.4 Coating and characterization of virus-AuNPs complexes ..............105 5.2.5 Proof-of-concept tests .....................................................................106 5.2.6 Statistical analysis ...........................................................................106 5.3 Results and discussion ..................................................................................107
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