Literature DB >> 28448032

A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation.

Christopher J A Warner1, Subrata Dutta1, Alejandro R Foley1, Jevgenij A Raskatov2.   

Abstract

Amyloidogenic peptides such as the Alzheimer's disease-implicated Amyloid beta (Aβ), can present a significant challenge when trying to obtain high purity material. Here we present a tailored HPLC purification protocol to produce high-purity amyloid beta 42 (Aβ42) and amyloid beta 40 (Aβ40) peptides. We have found that the combination of commercially available hydrophobic poly(styrene/divinylbenzene) stationary phase, polymer laboratory reverse phase - styrenedivinylbenzene (PLRP-S) under high pH conditions, enables the attainment of high purity (>95%) Aβ42 in a single chromatographic run. The purification is highly reproducible and can be amended to both semi-preparative and analytical conditions depending upon the amount of material wished to be purified. The protocol can also be applied to the Aβ40 peptide with identical success and without the need to alter the method.

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Year:  2017        PMID: 28448032      PMCID: PMC5564433          DOI: 10.3791/55482

Source DB:  PubMed          Journal:  J Vis Exp        ISSN: 1940-087X            Impact factor:   1.355


  13 in total

1.  Methodological and chemical factors affecting amyloid beta peptide amyloidogenicity.

Authors:  M G Zagorski; J Yang; H Shao; K Ma; H Zeng; A Hong
Journal:  Methods Enzymol       Date:  1999       Impact factor: 1.600

Review 2.  Alzheimer's disease.

Authors:  Henry W Querfurth; Frank M LaFerla
Journal:  N Engl J Med       Date:  2010-01-28       Impact factor: 91.245

3.  Use of reversed-phase high-performance liquid chromatography on polystyrene-divinylbenzene columns for the rapid separation and purification of acid-soluble nuclear proteins.

Authors:  N Z Zhelev; M J Barratt; L C Mahadevan
Journal:  J Chromatogr A       Date:  1997-02-28       Impact factor: 4.759

4.  On-line high-performance liquid chromatography/mass spectrometric investigation of amyloid-beta peptide variants found in Alzheimer's disease.

Authors:  A J Thompson; T K Lim; C J Barrow
Journal:  Rapid Commun Mass Spectrom       Date:  1999       Impact factor: 2.419

5.  An improved method of preparing the amyloid beta-protein for fibrillogenesis and neurotoxicity experiments.

Authors:  Y Fezoui; D M Hartley; J D Harper; R Khurana; D M Walsh; M M Condron; D J Selkoe; P T Lansbury; A L Fink; D B Teplow
Journal:  Amyloid       Date:  2000-09       Impact factor: 7.141

6.  Photo-induced cross-linking of unmodified proteins (PICUP) applied to amyloidogenic peptides.

Authors:  Farid Rahimi; Panchanan Maiti; Gal Bitan
Journal:  J Vis Exp       Date:  2009-01-12       Impact factor: 1.355

7.  Alzheimer's disease-affected brain: presence of oligomeric A beta ligands (ADDLs) suggests a molecular basis for reversible memory loss.

Authors:  Yuesong Gong; Lei Chang; Kirsten L Viola; Pascale N Lacor; Mary P Lambert; Caleb E Finch; Grant A Krafft; William L Klein
Journal:  Proc Natl Acad Sci U S A       Date:  2003-08-18       Impact factor: 11.205

8.  Amyloid beta -protein (Abeta) assembly: Abeta 40 and Abeta 42 oligomerize through distinct pathways.

Authors:  Gal Bitan; Marina D Kirkitadze; Aleksey Lomakin; Sabrina S Vollers; George B Benedek; David B Teplow
Journal:  Proc Natl Acad Sci U S A       Date:  2002-12-27       Impact factor: 11.205

9.  Mutations enhance the aggregation propensity of the Alzheimer's A beta peptide.

Authors:  Woojin Kim; Michael H Hecht
Journal:  J Mol Biol       Date:  2008-01-11       Impact factor: 5.469

10.  Sequence-engineered mRNA Without Chemical Nucleoside Modifications Enables an Effective Protein Therapy in Large Animals.

Authors:  Andreas Thess; Stefanie Grund; Barbara L Mui; Michael J Hope; Patrick Baumhof; Mariola Fotin-Mleczek; Thomas Schlake
Journal:  Mol Ther       Date:  2015-06-08       Impact factor: 11.454

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  2 in total

1.  A robust preparation method for the amyloidogenic and intrinsically disordered amyloid-α peptide.

Authors:  Ariel J Kuhn; Jevgenij A Raskatov
Journal:  J Pept Sci       Date:  2022-05-06       Impact factor: 2.408

2.  A crystal-structural study of Pauling-Corey rippled sheets.

Authors:  Ariel J Kuhn; Beatriz Ehlke; Timothy C Johnstone; Scott R J Oliver; Jevgenij A Raskatov
Journal:  Chem Sci       Date:  2021-12-08       Impact factor: 9.825

  2 in total

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