Influence of the polydispersity of pH 2 and pH 3.5 beta-lactoglobulin amyloid fibril solutions on analytical methods

Timon R. Heyn, Vasil M. Garamus, Hendrikje R. Neumann, Maximilian J. Uttinger, Tobias Guckeisen, Monique Heuer, Christine Selhuber-Unkel, Wolfgang Peukert, Julia K. Keppler*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)

Abstract

It is well known that amyloid beta-lactoglobulin (BLG) fibril solutions contain a heterogeneous mixture of amyloid aggregates and non-amyloid material. However, few information are available on how strongly separated fractions of different morphologies (straight fibrils at pH 2 and worm-like aggregates at pH 3.5) vary with respect to physicochemical properties and building blocks as most analyses are conducted with unfractionized solutions where superposition effects occur. The pH-value shift resulted in an altered degree of acid hydrolysis which led to dissimilar building blocks of the aggregates (peptides at pH 2, non-hydrolyzed protein at pH 3.5). The respective separated amyloid and non-amyloid fractions showed significantly different size (SAXS, SEC, AUC) and charge properties (Zeta potential) than the whole samples. Strong superposition effects were evident with common analyses such as FTIR, TRP fluorescence and Thioflavin-T. At the same time, structural differences of pH 2 and pH 3.5 aggregates could be presented more clearly.

Original languageEnglish
Article number109211
JournalEuropean Polymer Journal
Volume120
DOIs
Publication statusPublished - Nov 2019

Fingerprint

Lactoglobulins
Polydispersity
Amyloid
Zeta potential
Peptides
Hydrolysis
Fluorescence
worms
Proteins
Acids
peptides
hydrolysis
proteins
fluorescence
acids
shift

Keywords

  • Amyloid aggregates
  • AUC
  • Beta-lactoglobulin
  • Building-blocks
  • Fibrils
  • Polydispersity
  • SAXS
  • Superposition effects
  • Whey-protein
  • Worm-like fibrils

Cite this

Heyn, Timon R. ; Garamus, Vasil M. ; Neumann, Hendrikje R. ; Uttinger, Maximilian J. ; Guckeisen, Tobias ; Heuer, Monique ; Selhuber-Unkel, Christine ; Peukert, Wolfgang ; Keppler, Julia K. / Influence of the polydispersity of pH 2 and pH 3.5 beta-lactoglobulin amyloid fibril solutions on analytical methods. In: European Polymer Journal. 2019 ; Vol. 120.
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title = "Influence of the polydispersity of pH 2 and pH 3.5 beta-lactoglobulin amyloid fibril solutions on analytical methods",
abstract = "It is well known that amyloid beta-lactoglobulin (BLG) fibril solutions contain a heterogeneous mixture of amyloid aggregates and non-amyloid material. However, few information are available on how strongly separated fractions of different morphologies (straight fibrils at pH 2 and worm-like aggregates at pH 3.5) vary with respect to physicochemical properties and building blocks as most analyses are conducted with unfractionized solutions where superposition effects occur. The pH-value shift resulted in an altered degree of acid hydrolysis which led to dissimilar building blocks of the aggregates (peptides at pH 2, non-hydrolyzed protein at pH 3.5). The respective separated amyloid and non-amyloid fractions showed significantly different size (SAXS, SEC, AUC) and charge properties (Zeta potential) than the whole samples. Strong superposition effects were evident with common analyses such as FTIR, TRP fluorescence and Thioflavin-T. At the same time, structural differences of pH 2 and pH 3.5 aggregates could be presented more clearly.",
keywords = "Amyloid aggregates, AUC, Beta-lactoglobulin, Building-blocks, Fibrils, Polydispersity, SAXS, Superposition effects, Whey-protein, Worm-like fibrils",
author = "Heyn, {Timon R.} and Garamus, {Vasil M.} and Neumann, {Hendrikje R.} and Uttinger, {Maximilian J.} and Tobias Guckeisen and Monique Heuer and Christine Selhuber-Unkel and Wolfgang Peukert and Keppler, {Julia K.}",
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Heyn, TR, Garamus, VM, Neumann, HR, Uttinger, MJ, Guckeisen, T, Heuer, M, Selhuber-Unkel, C, Peukert, W & Keppler, JK 2019, 'Influence of the polydispersity of pH 2 and pH 3.5 beta-lactoglobulin amyloid fibril solutions on analytical methods', European Polymer Journal, vol. 120, 109211. https://doi.org/10.1016/j.eurpolymj.2019.08.038

Influence of the polydispersity of pH 2 and pH 3.5 beta-lactoglobulin amyloid fibril solutions on analytical methods. / Heyn, Timon R.; Garamus, Vasil M.; Neumann, Hendrikje R.; Uttinger, Maximilian J.; Guckeisen, Tobias; Heuer, Monique; Selhuber-Unkel, Christine; Peukert, Wolfgang; Keppler, Julia K.

In: European Polymer Journal, Vol. 120, 109211, 11.2019.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Influence of the polydispersity of pH 2 and pH 3.5 beta-lactoglobulin amyloid fibril solutions on analytical methods

AU - Heyn, Timon R.

AU - Garamus, Vasil M.

AU - Neumann, Hendrikje R.

AU - Uttinger, Maximilian J.

AU - Guckeisen, Tobias

AU - Heuer, Monique

AU - Selhuber-Unkel, Christine

AU - Peukert, Wolfgang

AU - Keppler, Julia K.

PY - 2019/11

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N2 - It is well known that amyloid beta-lactoglobulin (BLG) fibril solutions contain a heterogeneous mixture of amyloid aggregates and non-amyloid material. However, few information are available on how strongly separated fractions of different morphologies (straight fibrils at pH 2 and worm-like aggregates at pH 3.5) vary with respect to physicochemical properties and building blocks as most analyses are conducted with unfractionized solutions where superposition effects occur. The pH-value shift resulted in an altered degree of acid hydrolysis which led to dissimilar building blocks of the aggregates (peptides at pH 2, non-hydrolyzed protein at pH 3.5). The respective separated amyloid and non-amyloid fractions showed significantly different size (SAXS, SEC, AUC) and charge properties (Zeta potential) than the whole samples. Strong superposition effects were evident with common analyses such as FTIR, TRP fluorescence and Thioflavin-T. At the same time, structural differences of pH 2 and pH 3.5 aggregates could be presented more clearly.

AB - It is well known that amyloid beta-lactoglobulin (BLG) fibril solutions contain a heterogeneous mixture of amyloid aggregates and non-amyloid material. However, few information are available on how strongly separated fractions of different morphologies (straight fibrils at pH 2 and worm-like aggregates at pH 3.5) vary with respect to physicochemical properties and building blocks as most analyses are conducted with unfractionized solutions where superposition effects occur. The pH-value shift resulted in an altered degree of acid hydrolysis which led to dissimilar building blocks of the aggregates (peptides at pH 2, non-hydrolyzed protein at pH 3.5). The respective separated amyloid and non-amyloid fractions showed significantly different size (SAXS, SEC, AUC) and charge properties (Zeta potential) than the whole samples. Strong superposition effects were evident with common analyses such as FTIR, TRP fluorescence and Thioflavin-T. At the same time, structural differences of pH 2 and pH 3.5 aggregates could be presented more clearly.

KW - Amyloid aggregates

KW - AUC

KW - Beta-lactoglobulin

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KW - Fibrils

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KW - SAXS

KW - Superposition effects

KW - Whey-protein

KW - Worm-like fibrils

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DO - 10.1016/j.eurpolymj.2019.08.038

M3 - Article

VL - 120

JO - European Polymer Journal

JF - European Polymer Journal

SN - 0014-3057

M1 - 109211

ER -