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, M.; Selhuber‐Unkel, Christine; Peukert, Wolfgang; Keppler, Julia K.

doi:10.1016/j.eurpolymj.2019.08.038

Cited by 39 publications

(13 citation statements)

References 58 publications

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“…WPI fibril formation (shown schematically in Figure 1 a) was influenced by pH, as measurements of fibrillar yield at pH 2 (approximately 25%) and pH 3.5 (>40%) demonstrated ( Figure 1 b). Similar observations were reported previously [ 5 , 6 , 7 ]. Differences in yield are attributed to differences in the fibril building blocks, which are specific acid hydrolyzed peptides at pH 2 [ 4 ], but unspecific non-hydrolyzed proteins at pH 3.5 [ 5 ].…”

Section: Resultssupporting

confidence: 93%

“…SEM images confirmed that fibril coatings withstood washing and drying. Fibrils prepared at pH 3.5 ( Figure 2 c) appeared to be shorter and less straight than those formed at pH 2 ( Figure 2 b), in agreement with previous studies [ 5 ]. Fibrils were detected by SEM after autoclaving ( Supplementary Information, Figure S1 ); hence, they withstand sterilization.…”

Section: Resultssupporting

confidence: 91%

“…Substrates were rinsed with Milli-Q to remove non-adhered fibrils, air-dried and autoclaved (121 • C, 15 min). SEM, CA measurements and fibrillar yield quantification were performed as described previously [5,7,31,32].…”

Section: Methodsmentioning

confidence: 99%

“…Upon heating for several hours under acidic conditions (<pH 3), β-lactoglobulin degrades into smaller peptides, which undergo self-assembly to form amyloid fibrils several micrometers long and a few nm thick [4]. Heating at pH 3.5 leads to worm-like structures which consist of whole protein instead of peptides [5][6][7].…”

Section: Introductionmentioning

confidence: 99%

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Dairy-Inspired Coatings for Bone Implants from Whey Protein Isolate-Derived Self-Assembled Fibrils

Rabe

Hempel

Martocq

et al. 2020

IJMS

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To improve the integration of a biomaterial with surrounding tissue, its surface properties may be modified by adsorption of biomacromolecules, e.g., fibrils. Whey protein isolate (WPI), a dairy industry by-product, supports osteoblastic cell growth. WPI’s main component, β-lactoglobulin, forms fibrils in acidic solutions. In this study, aiming to develop coatings for biomaterials for bone contact, substrates were coated with WPI fibrils obtained at pH 2 or 3.5. Importantly, WPI fibrils coatings withstood autoclave sterilization and appeared to promote spreading and differentiation of human bone marrow stromal cells (hBMSC). In the future, WPI fibrils coatings could facilitate immobilization of biomolecules with growth stimulating or antimicrobial properties.

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Section: Resultssupporting

confidence: 93%

Section: Resultssupporting

confidence: 91%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Dairy-Inspired Coatings for Bone Implants from Whey Protein Isolate-Derived Self-Assembled Fibrils

Rabe

Hempel

Martocq

et al. 2020

IJMS

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“…Since the conversion rate is rather low, approximately 30–40% of the proteins are converted to fibrils. The residual populations consist of monomers, oligomers and random aggregates and, thus, the samples show a high polydispersity (Heyn et al 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Measurement of length distribution of beta-lactoglobulin fibrils by multiwavelength analytical ultracentrifugation

et al. 2020

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The whey protein beta-lactoglobulin is the building block of amyloid fibrils which exhibit a great potential in various applications. These include stabilization of gels or emulsions. During biotechnological processing, high shear forces lead to fragmentation of fibrils and therefore to smaller fibril lengths. To provide insight into such processes, pure straight amyloid fibril dispersions (prepared at pH 2) were produced and sheared using the rotor stator setup of an Ultra Turrax. In the first part of this work, the sedimentation properties of fragmented amyloid fibrils sheared at different stress levels were analyzed with mulitwavelength analytical ultracentrifugation (AUC). Sedimentation data analysis was carried out with the boundary condition that fragmented fibrils were of cylindrical shape, for which frictional properties are known. These results were compared with complementary atomic force microscopy (AFM) measurements. We demonstrate how the sedimentation coefficient distribution from AUC experiments is influenced by the underlying length and diameter distribution of amyloid fibrils. In the second part of this work, we show how to correlate the fibril size reduction kinetics with the applied rotor revolution and the resulting energy density, respectively, using modal values of the sedimentation coefficients obtained from AUC. Remarkably, the determined scaling laws for the size reduction are in agreement with the results for other material systems, such as emulsification processes or the size reduction of graphene oxide sheets.

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New Prospects for Particle Characterization Using Analytical Centrifugation with Sector‐Shaped Centerpieces

Uttinger

Boldt²,

Wawra

et al. 2020

Part & Part Syst Charact

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Analytical centrifugation (AC) has recently shown great potential for the accurate determination of particle size distributions. The well‐established LUMiSizer(R) is customized by a new design allowing for higher rotor speeds, improved thermal insulation, and measurement cell assembly. The latter enables sedimentation analysis of nanoparticles (NPs) in sector‐shaped centerpieces. The measurement window of AC experiments is assessed by the Peclet (Pe) number. It is shown that at low Pe numbers (0.7 < Pe < 30), sedimentation, and diffusion can be accurately and simultaneously analyzed from the sedimentation boundaries within one experiment. Moreover, sedimentation properties can be reliably determined up to Pe numbers of 4000. The thermal characteristic throughout the sedimentation analysis is validated by measuring polystyrene particles at elevated temperatures. Moreover, the performance of the setup is demonstrated by determining the sedimentation properties of SiO2 NPs at intermediate Pe numbers in excellent agreement with results from analytical ultracentrifugation experiments. Finally, for the first time, an accurate analysis of the core–shell properties of Au NPs via AC is presented. By combining the analysis of sedimentation and diffusional properties at low Pe numbers, the shell thickness of the stabilizer cetyltrimethylammonium bromide alongside the core diameter distribution of the Au NPs is determined.

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

Cited by 39 publications

References 58 publications

Dairy-Inspired Coatings for Bone Implants from Whey Protein Isolate-Derived Self-Assembled Fibrils

Dairy-Inspired Coatings for Bone Implants from Whey Protein Isolate-Derived Self-Assembled Fibrils

Measurement of length distribution of beta-lactoglobulin fibrils by multiwavelength analytical ultracentrifugation

New Prospects for Particle Characterization Using Analytical Centrifugation with Sector‐Shaped Centerpieces

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