Analysis of Aβ (1-40) and Aβ (1-42) monomer and fibrils by capillary electrophoresis

Picou, Ryan A.; Kheterpal, Indu; Wellman, Amber D.; Minnamreddy, Madhavi; Ku, Ginger; Gilman, S. Douglass

doi:10.1016/j.jchromb.2011.01.030

Cited by 27 publications

(28 citation statements)

References 34 publications

(64 reference statements)

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“…Several sharp peaks (peak clusters) differing in number and electrophoretic mobility have also been observed by Picou et al. when studying aggregation of beta‐amyloid, also proteins, in CE. Similarly, Haugg et al.…”

Section: Resultssupporting

confidence: 62%

Affinity capillary electrophoresis to evaluate the complex formation between poliovirus and nanobodies

Halewyck

Schotte

Oita

et al. 2014

J of Separation Science

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It was demonstrated that nanobodies with an in vitro neutralizing activity against poliovirus type 1 interact with native virions. Here, the use of capillary electrophoresis was investigated as an alternative technique for the evaluation of the formation of nanobody-poliovirus complexes, and therefore predicting the in vitro neutralizing activity of the nanobodies. The macromolecules are preincubated offline in a specific nanobody-to-virus ratio and analyzed by capillary electrophoresis with UV detection. At low nanobody-to-virus ratios, a clear shift in migration time of the viral peak was observed. A broad peak was obtained, indicating the presence of a heterogeneous population of nanobody-virion complexes, caused by the binding of different numbers of nanobodies to the virus particle. At elevated nanobody-to-virus ratios, a cluster of peaks appeared, showing an additional increase in migration times. It was shown that, at these high molar excesses, aggregates were formed. The developed capillary electrophoresis method can be used as a rapid, qualitative screening for the affinity between poliovirus and nanobodies, based on a clearly visible and measurable shift in migration time. The advantages of this technique include that there is no need for antigen immobilization as in enzyme-linked immunosorbent assays or surface plasmon resonance for the use of radiolabeled virus or for the performance of labor- and time-intensive plaque-forming neutralization assays.

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Section: Resultssupporting

confidence: 62%

Affinity capillary electrophoresis to evaluate the complex formation between poliovirus and nanobodies

Halewyck

Schotte

Oita

et al. 2014

J of Separation Science

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“…As compared to previous CE-UV [23] or CE-LIF [24,25] studies that did not consider blanks (i.e., incubated or un-incubated matrix), the present work has revealed factors leading to interfering spikes, i.e., spikes related to material other than Aβ aggregates. Clearly, important causes of interfering spikes may be silica particles released from the capillary inlet in response to the mechanical stress to capillary occurring during the injection step and/or tiny dusts ripped out from the rubber stopper of vials.…”

Section: Discussionmentioning

confidence: 47%

“…As it was not possible to have access to an electronic microscopy platform when performing the present study, the presence of fibrillar aggregates was not confirmed using an imaging approach. However, one can safely assume that our samples contained fibrils of Aβ peptides, because the incubation conditions we used (buffer, temperature, duration) are well known to lead to Aβ aggregates, some of them being fibrils [15,[23][24][25].…”

Section: Discussionmentioning

confidence: 99%

“…CE with UV detection was employed for the analysis of several Aβ monomers [16,17], for the separation of monomers and transient soluble oligomers of Aβ 1-40 or 1-42 using neutral saline background electrolyte [18][19][20][21] or a polymeric separation matrix [22]. Moreover, CE-UV allowed the separation of monomers, large aggregates (migrating as wide peak) and fibrils (migrating as sharp peaks or spikes) [23]. In order to improve sensitivity and selectivity, CE-LIF was also employed for the analysis of fibrils of Aβ.…”

Section: Introductionmentioning

confidence: 99%

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Advances and Pitfalls in the Capillary Electrophoresis Analysis of Aggregates of Beta Amyloid Peptides

Denoroy

Parrot

2017

Separations

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Alzheimer's disease is characterized by the accumulation of brain amyloid plaques composed of aggregates of amyloid β (Aβ) peptides. The present paper describes a novel and easy-to-run capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) method for the specific analysis of fibrillar forms of Aβ aggregates obtained after in vitro incubation of Aβ 1-40 monomer. For that purpose, an affinity CE-LIF approach in which the ligand thioflavine T was added to the running buffer has been used, leading to the separation and detection of various fibrillar aggregates which migrated as spikes. The procedure has been optimized to get spikes only corresponding to Aβ aggregates, through the careful elimination of interfering factors and the electrophoretic validation of the link between spikes and particulate material. This method exhibited semi-quantification capabilities, led to the separation of Aβ fibrillar aggregates of different sizes and showed that highly concentrated solutions of Aβ peptides led to the formation of aggregates of larger size than lower-concentrated solution did. Advances brought by this method as well as future development needed to overcome its present limitations are discussed.

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“…30 Picou et al used UV-CE to separate Ab 1-42 monomer from fibrils. 31 Kato et al used CE with laser induced fluorescence (LIF) to separate two different Ab 1-42 aggregates. 32 Brambilla et al used LIF-CE to show that Ab aggregation is altered in the presence of PEGylated nanoparticles.…”

Section: Introductionmentioning

confidence: 99%

Field amplified sample stacking of amyloid beta (1-42) oligomers using capillary electrophoresis

Paracha

Hestekin

2016

Biomicrofluidics

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Oligomeric forms of the amyloid beta (Aβ) protein have been indicated to be an important factor in the development of Alzheimer's disease (AD). Since the oligomeric forms of Aβ can vary in size and conformation, it is vital to understand the early stages of Aβ aggregation in order to improve the care and treatment of patients with AD. This is the first study to determine the effect of field amplified sample stacking (FASS) on the separation of oligomeric forms of Aβ1-42 using capillary electrophoresis (CE) with ultraviolet (UV) detection. UV-CE was able to separate two different species of Aβ1-42 oligomers (<7 mers and 7–22 mers). Although FASS required the use of a higher ionic strength buffer, Aβ1-42 oligomers had the same aggregation behavior as under the non-FASS conditions with only small changes in the amounts of oligomers observed. In general, FASS provided smaller peak widths (>75% average reduction) and increased peak heights (>60% average increase) when compared to non-FASS conditions. UV-CE with FASS also provided higher resolution between the Aβ1-42 oligomers for all aggregation time points studied. In addition, Congo red and Orange G inhibition studies were used to help evaluate the conformation of the observed species. This work demonstrates the ability of UV-CE employing FASS to provide higher resolution between oligomeric forms of Aβ1-42 without significantly altering their aggregation.

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Analysis of Aβ (1-40) and Aβ (1-42) monomer and fibrils by capillary electrophoresis

Cited by 27 publications

References 34 publications

Affinity capillary electrophoresis to evaluate the complex formation between poliovirus and nanobodies

Affinity capillary electrophoresis to evaluate the complex formation between poliovirus and nanobodies

Advances and Pitfalls in the Capillary Electrophoresis Analysis of Aggregates of Beta Amyloid Peptides

Field amplified sample stacking of amyloid beta (1-42) oligomers using capillary electrophoresis

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