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  Lyophilization induces physicochemical alterations in cryptococcal exopolysaccharide

Wear, M. P., Hargett, A. A., Kelly, J. E., McConnell, S. A., Crawford, C., Freedberg, D. I., et al. (2022). Lyophilization induces physicochemical alterations in cryptococcal exopolysaccharide. Carbohydrate Polymers, 291: 119547. doi:10.1016/j.carbpol.2022.119547.

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 Creators:
Wear, Maggie P., Author
Hargett, Audra A., Author
Kelly, John E., Author
McConnell, Scott A., Author
Crawford, Conor1, Author              
Freedberg, Darón I., Author
Stark, Ruth E., Author
Casadevall, Arturo, Author
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1External Organizations, ou_persistent22              

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Free keywords: Cryptococcus; NMR; DLS; TEM; Exopolysaccharide; Lyophilization
 Abstract: Microbial polysaccharide characterization requires purification that often involves detergent precipitation and lyophilization. Here we examined physicochemical changes following lyophilization of Cryptococcus neoformans exopolysaccharide (EPS). Solution 1H Nuclear Magnetic Resonance (NMR) reveals significant anomeric signal attenuation following lyophilization of native EPS while 1H solid-state Nuclear Magnetic Resonance (ssNMR) shows few changes, suggesting diminished molecular motion and consequent broadening of 1H NMR polysaccharide resonances. 13C ssNMR, dynamic light scattering, and transmission electron microscopy show that, while native EPS has rigid molecular characteristics and contains small, loosely packed polysaccharide assemblies, lyophilized and resuspended EPS is disordered and contains larger dense aggregates, suggesting that structural water molecules in the interior of the polysaccharide assemblies are removed during extensive lyophilization. Importantly, mAbs to C. neoformans polysaccharide bind native EPS more strongly than lyophilized EPS. Together, these observations argue for caution when interpreting the biological and immunological attributes of polysaccharides that have been lyophilized to dryness.

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Language(s): eng - English
 Dates: 2022-04-292022
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: DOI: 10.1016/j.carbpol.2022.119547
 Degree: -

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Title: Carbohydrate Polymers
Source Genre: Journal
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Publ. Info: London [Eng.] : Elsevier
Pages: - Volume / Issue: 291 Sequence Number: 119547 Start / End Page: - Identifier: ISSN: 0144-8617