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Hauptverfasser: Bal, Vivekananda, Wolfrum, Jackie M., Barone, Paul W., Springs, Stacy L., Sinskey, Anthony J., Kotin, Robert M., Braatz, Richard D.
Format: Preprint
Veröffentlicht: 2025
Schlagworte:
Online-Zugang:https://arxiv.org/abs/2510.05867
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author Bal, Vivekananda
Wolfrum, Jackie M.
Barone, Paul W.
Springs, Stacy L.
Sinskey, Anthony J.
Kotin, Robert M.
Braatz, Richard D.
author_facet Bal, Vivekananda
Wolfrum, Jackie M.
Barone, Paul W.
Springs, Stacy L.
Sinskey, Anthony J.
Kotin, Robert M.
Braatz, Richard D.
contents Physicochemical characterization of materials is central to the field of science and engineering and is essential to design new/engineered materials with specific properties. Assays available for small-molecules, e.g., XRD, NMR, LC-MS, can't be applied to macromolecules easily. Thus, it is extremely challenging to characterize complex materials such as adeno-associated virus capsids (MW 5.8MDa). Capsid crystals produced in hanging-drop are characterized in-situ using cross-polarized light and ex-situ using scanning electron microscopy, energy dispersive X-ray, and transmission electron microscopy. Cross-polarized light can be used to identify capsid crystals within a heterogenous-system of kosmotropic/chaotropic-salt crystals, fibers, dense solid-phase, opaques crystals. Despite highly-conserved structures, crystal birefringence suggests that capsids possess serotype-specific structural differences. SEM demonstrated that crystal-growth occurs by random 2D-nucleation followed by kink-site attachment and/or spread by more 2D-nuclei and proteinaceous assemblies tend to form semi-crystalline solids appearing as dense/opaque materials. EDX shows that C, O, and N are present in ratio of 2.33+-0.222:1:0.583+-0.019 for serotypes 5, 8, and 9 and can be an alternative to protein sequencing-based virus identification. Biological macromolecular assemblies are found to facilitate plural-scattering responsible for Kikuchi-diffraction pattern even for thin-crystals (~300nm). For an optimal spot-diffraction, crystals must possess at least one dimension consisting of at most 8 layers of capsids.
format Preprint
id arxiv_https___arxiv_org_abs_2510_05867
institution arXiv
publishDate 2025
record_format arxiv
spellingShingle Advanced Characterization Methods for Crystals of an Extraordinary Material
Bal, Vivekananda
Wolfrum, Jackie M.
Barone, Paul W.
Springs, Stacy L.
Sinskey, Anthony J.
Kotin, Robert M.
Braatz, Richard D.
Chemical Physics
Physicochemical characterization of materials is central to the field of science and engineering and is essential to design new/engineered materials with specific properties. Assays available for small-molecules, e.g., XRD, NMR, LC-MS, can't be applied to macromolecules easily. Thus, it is extremely challenging to characterize complex materials such as adeno-associated virus capsids (MW 5.8MDa). Capsid crystals produced in hanging-drop are characterized in-situ using cross-polarized light and ex-situ using scanning electron microscopy, energy dispersive X-ray, and transmission electron microscopy. Cross-polarized light can be used to identify capsid crystals within a heterogenous-system of kosmotropic/chaotropic-salt crystals, fibers, dense solid-phase, opaques crystals. Despite highly-conserved structures, crystal birefringence suggests that capsids possess serotype-specific structural differences. SEM demonstrated that crystal-growth occurs by random 2D-nucleation followed by kink-site attachment and/or spread by more 2D-nuclei and proteinaceous assemblies tend to form semi-crystalline solids appearing as dense/opaque materials. EDX shows that C, O, and N are present in ratio of 2.33+-0.222:1:0.583+-0.019 for serotypes 5, 8, and 9 and can be an alternative to protein sequencing-based virus identification. Biological macromolecular assemblies are found to facilitate plural-scattering responsible for Kikuchi-diffraction pattern even for thin-crystals (~300nm). For an optimal spot-diffraction, crystals must possess at least one dimension consisting of at most 8 layers of capsids.
title Advanced Characterization Methods for Crystals of an Extraordinary Material
topic Chemical Physics
url https://arxiv.org/abs/2510.05867