Isoform Separation by a Mixed-mode Resin, TOYOPEARL MX-Trp-650M

Author(s): Tsutomu Arakawa*.

Journal Name: Current Protein & Peptide Science

Volume 20 , Issue 1 , 2019

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Abstract:

TOYOPEARL particles are cross-linked hydroxylated methacrylic polymers available in different pore and particle sizes. They are conjugated with different ligands to generate ion-exchange, hydrophobic interaction and affinity resins. They have excellent physical and chemical properties. A mixed-mode resin, TOYOPEARL MX-Trp-650M, is made of this particle with tryptophan conjugated via N-terminal amino group and hence has both hydrophobic/aromatic side chain and carboxyl group. In this review, I will summarize the properties of the TOYOPEARL particles and MX-Trp-650M resin and application of this resin for purification of proteins and in some detail the separation of disulfide (SS)- scrambled oligomers of insulin-like growth factor-1 (IGF-1). For this particular application, the intact IGF-1 was used to examine binding and elution conditions of TOYOSCREEN MX-Trp-650M column. Strong binding was obtained at pH 4.0, at which arginine, but not NaCl, resulted in elution. Both NaCl and arginine resulted in elution at pH 6.5. In addition, a pH gradient from 4.0 to 8.5 was effective. When applied to SS-scrambled IGF-1 oligomers, both pH and arginine gradient exhibited an efficient separation of the oligomers.

Keywords: Mixed-mode chromatography, TOYOPEARL MX-Trp-650M, arginine, isoform, aggregates, polymers.

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Article Details

VOLUME: 20
ISSUE: 1
Year: 2019
Page: [61 - 64]
Pages: 4
DOI: 10.2174/1389203718666171009111355
Price: $58

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