scholarly journals Measuring molecular interactions in solution using multi-wavelength analytical ultracentrifugation: combining spectral analysis with hydrodynamics

2019 ◽  
Vol 41 (2) ◽  
pp. 14-18 ◽  
Author(s):  
Borries Demeler
Keyword(s):  
Analytical Ultracentrifugation ◽  
Heterogeneous Systems ◽  
Colloidal Properties ◽  
Resolution Capability ◽  
Technological Advances ◽  
Data Density ◽  
Hydrodynamic Separation ◽  
Multi Wavelength ◽  
Spectral Absorbance ◽  
Indispensable Tool

In 1926, the Swedish scientist Theodor Svedberg was awarded the Nobel Prize in Chemistry for his work on a disperse system, and for studying the colloidal properties of proteins. This work was, to a large extent, made possible by his invention of a revolutionary tool, the analytical ultracentrifuge. These days, technological advances in hardware and computing have transformed the field of analytical ultracentrifugation (AUC) by enabling entirely new classes of experiments and modes of measurement unimaginable by Svedberg, making AUC once again an indispensable tool for modern biomedical research. In this article these advances and their impact on studies of interacting molecules will be discussed, with particular emphasis on a new method termed multi-wavelength analytical ultracentrifugation (MWL-AUC). Novel detectors allow us to add a second dimension to the separation of disperse and heterogeneous systems: in addition to the traditional hydrodynamic separation of colloidal mixtures, it is now possible to identify the sedimenting molecules by their spectral absorbance properties. The potential for this advance is significant for the study of a large range of systems. A further advance has occurred in data management and computational capabilities, opening doors to improved analysis methods, as well as direct networking with the instrument, facilitating data acquisition and data handling, and significant increases in data density from faster detectors with higher resolution capability.

A Study on the Multi-Wavelength Drop Analysis Technology

2005 ◽  
Vol 295-296 ◽  
pp. 215-220 ◽  
Author(s):  
G. Hao ◽  
Zu Rong Qiu ◽  
Guo Xiong Zhang ◽  
A.P. Zhang
Keyword(s):  
Light Intensity ◽  
Liquid Drop ◽  
Instrumental Method ◽  
Test Results ◽  
Light Wavelength ◽  
Drop Volume ◽  
Signal Process ◽  
Key Technology ◽  
Resolution Capability ◽  
Multi Wavelength

An instrumental method for measuring the properties of liquids has been developed. The instrument, called the fiber-capacitive drop analyzer (FCDA), is based on a specially designed drop sensor by means of the fiber drop analyzing technology and the capacitive drop analyzing technology, and produces a volume-based fiber drop trace (VFDT), which shows the relation between the light intensity passing through the liquid drop and the instant drop volume. Preliminary experiments have been carried out and these prove that VFDT externalizes the overall properties of tested liquids and is suitable for fine discrimination among different kinds of liquids. In this paper, a multi-wavelength drop analysis (MWDA) method is described. In this case, the light source of the FCDA is developed from a single wavelength to a multi-wavelength. A special filter is employed to sort out the signals from the mixed multi-wavelength signals and it becomes possible to get two or three VFDT with different light wavelength at the same time. As one knows, a material will absorb the light on certain wavelengths. It means that, for a mixed liquid, one can get different VFDTs by MWDA at the same time. By this way, the resolution capability of a FCDA is enhanced very much due to more information including in those VFDTs. A special signal process circuit, the key technology of the MWDA, is described in this paper. The test results show that the new circuit had satisfied the MWDA method well and will be very useful to an instrument of the FCDA with MWDA.

scholarly journals Multi-Wavelength Analytical Ultracentrifugation of Biopolymer Mixtures and Interactions

2021 ◽  
Author(s):  
Borries Demeler ◽  
Amy Henrickson ◽  
Gary Gorbet ◽  
Alexey Savelyev ◽  
Minji Kim ◽  
...  
Keyword(s):  
Small Molecules ◽  
Analytical Ultracentrifugation ◽  
Optical Systems ◽  
Hydrodynamic Properties ◽  
Advantages And Disadvantages ◽  
Design Considerations ◽  
Free Dna ◽  
Wide Range ◽  
Analysis Strategies ◽  
Multi Wavelength

Abstract Multi-wavelength analytical ultracentrifugation (MW-AUC) is a recent development made possible by new analytical ultracentrifuge optical systems. MW-AUC is suitable for a wide range of applications and biopolymer systems and is poised to become an essential tool to characterize macromolecular interactions. It adds an orthogonal spectral dimension to the traditional hydrodynamic characterization by exploiting unique chromophores in analyte mixtures that may or may not interact. Here we illustrate the utility of MW-AUC for representative classes of challenging biopolymer systems, including interactions between mixtures of different sized proteins with small molecules, mixtures of loaded and empty viral AAV capsids contaminated with free DNA, and mixtures of different proteins, where some have identical hydrodynamic properties, all of which are difficult to resolve with traditional AUC methods. We explain the improvement in resolution and information content obtained by this technique compared to traditional single- or dual-wavelength approaches. We discuss experimental design considerations and limitations of the method, and address the advantages and disadvantages of the two MW optical systems available today, and the differences in data analysis strategies between the two systems.

Multi-wavelength analytical ultracentrifugation as a tool to characterise protein–DNA interactions in solution

2020 ◽  
Vol 49 (8) ◽  
pp. 819-827 ◽  
Author(s):  
Christopher R. Horne ◽  
Amy Henrickson ◽  
Borries Demeler ◽  
Renwick C. J. Dobson
Keyword(s):  
Analytical Ultracentrifugation ◽  
Dna Interactions ◽  
Protein Dna Interactions ◽  
Multi Wavelength

scholarly journals Consensus model of a cyanobacterial light-dependent protochlorophyllide oxidoreductase in its pigment-free apo-form and photoactive ternary complex

2019 ◽  
Vol 2 (1) ◽  
Author(s):  
Judith Schneidewind ◽  
Frank Krause ◽  
Marco Bocola ◽  
Andreas Maximilian Stadler ◽  
Mehdi D. Davari ◽  
...  
Keyword(s):  
Ternary Complex ◽  
Analytical Ultracentrifugation ◽  
Structural Information ◽  
Md Simulations ◽  
Consensus Model ◽  
Photosynthetic Organisms ◽  
X Ray Scattering ◽  
Multi Wavelength ◽  
Modelling Studies ◽  
Chlorophyll Precursor

Abstract Photosynthetic organisms employ two different enzymes for the reduction of the C17 = C18 double bond of protochlorophyllide (Pchlide), yielding the chlorophyll precursor chlorophyllide. First, a nitrogenase-like, light-independent (dark-operative) Pchlide oxidoreductase and secondly, a light-dependent Pchlide oxidoreductase (LPOR). For the latter enzyme, despite decades of research, no structural information is available. Here, we use protein structure modelling, molecular dynamics (MD) simulations combined with multi-wavelength analytical ultracentrifugation (MWA-AUC) and small angle X-ray scattering (SAXS) experiments to derive a consensus model of the LPOR apoprotein and the substrate/cofactor/LPOR ternary complex. MWA-AUC and SAXS experiments independently demonstrate that the apoprotein is monomeric, while ternary complex formation induces dimerization. SAXS-guided modelling studies provide a full-length model of the apoprotein and suggest a tentative mode of dimerization for the LPOR ternary complex, supported by published cross-link constraints. Our study provides a first impression of the LPOR structural organization.

scholarly journals Multi‐wavelength Analytical Ultracentrifugation of Human Serum Albumin complexed with Porphyrin

2018 ◽  
Vol 32 (S1) ◽  
Author(s):  
Courtney Nicole Johnson ◽  
Gary E. Gorbet ◽  
Heidi Ramsower ◽  
Julio Uriquidi ◽  
Lorenzo Brancaleon ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Analytical Ultracentrifugation ◽  
Multi Wavelength

Density Matching Multi-wavelength Analytical Ultracentrifugation to Measure Drug Loading of Lipid Nanoparticle Formulations

ACS Nano ◽  
2021 ◽  
Author(s):  
Amy Henrickson ◽  
Jayesh A. Kulkarni ◽  
Josh Zaifman ◽  
Gary E. Gorbet ◽  
Pieter R. Cullis ◽  
...  
Keyword(s):  
Analytical Ultracentrifugation ◽  
Drug Loading ◽  
Lipid Nanoparticle ◽  
Multi Wavelength

scholarly journals Multi-Wavelength Analytical Ultracentrifugation of Biopolymer Mixtures and Interactions

2021 ◽  
Author(s):  
Amy Henrickson ◽  
Gary E Gorbet ◽  
Alexey Savelyev ◽  
Minji Kim ◽  
Sarah K Schultz ◽  
...  
Keyword(s):  
Small Molecules ◽  
Analytical Ultracentrifugation ◽  
Optical Systems ◽  
Hydrodynamic Properties ◽  
Advantages And Disadvantages ◽  
Design Considerations ◽  
Free Dna ◽  
Wide Range ◽  
Analysis Strategies ◽  
Multi Wavelength

Multi-wavelength analytical ultracentrifugation (MW-AUC) is a recent development made possible by new analytical ultracentrifuge optical systems. MW-AUC is suitable for a wide range of applications and biopolymer systems and is poised to become an essential tool to characterize macromolecular interactions. It adds an orthogonal spectral dimension to the traditional hydrodynamic characterization by exploiting unique chromophores in analyte mixtures that may or may not interact. Here we illustrate the utility of MW-AUC for representative classes of challenging biopolymer systems, including interactions between mixtures of different sized proteins with small molecules, mixtures of loaded and empty viral AAV capsids contaminated with free DNA, and mixtures of different proteins, where some have identical hydrodynamic properties, all of which are difficult to resolve with traditional AUC methods. We explain the improvement in resolution and information content obtained by this technique compared to traditional single- or dual-wavelength approaches. We discuss experimental design considerations and limitations of the method, and address the advantages and disadvantages of the two MW optical systems available today, and the differences in data analysis strategies between the two systems.

scholarly journals 3D Land Administration: A Review and a Future Vision in the Context of the Spatial Development Lifecycle

2020 ◽  
Vol 9 (2) ◽  
pp. 107 ◽  
Author(s):  
Eftychia Kalogianni ◽  
Peter van Oosterom ◽  
Efi Dimopoulou ◽  
Christiaan Lemmen
Keyword(s):  
Heterogeneous Systems ◽  
Spatial Development ◽  
Domain Model ◽  
Data Reuse ◽  
Land Administration ◽  
3D Data ◽  
Development Lifecycle ◽  
Technological Advances ◽  
Sectoral Approach ◽  
Building Information

Land Administration practices worldwide rely mainly on 2D-based systems to define legal and other spatial boundaries related to land interests. However, the built environment is increasingly becoming spatially complex. Land administrators are challenged by an unprecedented demand to utilise space above and below earth’s surface. The relationships between people and land in vertical space can no longer be unambiguously represented in 2D. In addition, the current societal demand for sustainability in a collaborative environment and a lifecycle-thinking, is driving the need to integrate independent systems with standalone databases and methodologies, associated with different aspects of the Spatial Development lifeCycle (SDC). Land Administration Systems (LASs) are an important component of the SDC. Today, a LAS is often mandated and managed as a domain in isolation. Interaction and data reuse with the other phases of the SDC is limited and far from optimal. It is expected that effective 3D data collaboration, sharing, and reuse across the sectors and disciplines in the lifecycle will enable new ways of data harmonisation and use in this complex environment; will improve efficiency of design and data acquisition, as well as data quality (in relation to specific regulations); and will minimise inconsistencies and data loss within information flows. Overall, a cross-sectoral approach is directed towards improving the current state of the Land Administration (LA) domain. This paper consists of two parts. In the first, a review of the current situation, with respect to LASs is presented, concluding the needs for improvement in terms of effectiveness and consistency. In the second part, the vision for the future of LASs is introduced in a wider context, and as an important phase in the SDC, with regards to legal, technical, and organisational aspects. In this part, the needs and considerations that result from the evolving environment and the emerging technological advances are addressed, with a view to discussing a cross-sector approach to collect, maintain, reuse, and share 3D data. In such a cross-sectoral approach, various interoperability issues appear, making it necessary to introduce and use standards. In this respect, the ISO 19152:2012 Land Administration Domain Model (LADM) in its current Edition I, as well as in Edition II (expected in 2022) may serve as the standardised core structure of a 3D LAS, with respect to its role as further presented in this paper. In parallel, the evolution of the Building Information Modelling (BIM) in the design and construction industry, as well as the fact that BIM plays a central role in the life cycle of development projects, are well recognized. Emphasis is given on feasible reuse of BIM/IFC (Industry Foundation Class) data in a 3D LAS. Those considerations are addressed through a web-based system architecture for a future 3D LAS, thereby attempting to integrate heterogeneous systems in the SDC.

scholarly journals Multi-wavelength analytical ultracentrifugation: human serum albumin complexed with porphyrin identification

2018 ◽  
Vol 1 (1) ◽  
pp. 134-136
Author(s):  
C.N. Johnson ◽  
◽  
H.Х. Ramsower ◽  
J. Urquidi ◽  
L. Brancaleon ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Analytical Ultracentrifugation ◽  
Multi Wavelength

scholarly journals Multi-wavelength analytical ultracentrifugation of human serum albumin complexed with porphyrin

2018 ◽  
Vol 47 (7) ◽  
pp. 789-797 ◽  
Author(s):  
Courtney N. Johnson ◽  
Gary E. Gorbet ◽  
Heidi Ramsower ◽  
Julio Urquidi ◽  
Lorenzo Brancaleon ◽  
...  
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Analytical Ultracentrifugation ◽  
Multi Wavelength
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