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

[ Vol. 20 , Issue. 1 ]

Author(s):

Tsutomu Arakawa*   Pages 61 - 64 ( 4 )

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.

Affiliation:

Alliance Protein Laboratories, A Division of KBI Biopharma, 6042 Cornerstone Court West, San Diego, CA 92121

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