Aspartic acid incorporated monolithic columns for affinity glycoprotein purification

ARMUTCU ÇORMAN C., BERELİ N., Bayram E., UZUN L., Say R., DENİZLİ A.

COLLOIDS AND SURFACES B-BIOINTERFACES, vol.114, pp.67-74, 2014 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 114
  • Publication Date: 2014
  • Doi Number: 10.1016/j.colsurfb.2013.08.008
  • Journal Name: COLLOIDS AND SURFACES B-BIOINTERFACES
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.67-74
  • Keywords: Monolithic columns, IgG separation, Pseudospecific ligands, High performance liquid chromatography, POLY(GLYCIDYL METHACRYLATE) BEADS, PERFORMANCE LIQUID-CHROMATOGRAPHY, ANTIBODY PURIFICATION, ION-EXCHANGE, AMINO-ACIDS, SEPARATION, MEMBRANES, CRYOGELS, PROTEINS
  • Anadolu University Affiliated: Yes

Abstract

Novel aspartic acid incorporated monolithic columns were prepared to efficiently affinity purify immunoglobulin G (IgG) from human plasma. The monolithic columns were synthesised in a stainless steel HPLC column (20 cm x 5 mm id) by in situ bulk polymerisation of N-methacryloyl-L-aspartic acid (MAAsp), a polymerisable derivative of L-aspartic acid, and 2-hydroxyethyl methacrylate (HEMA). Monolithic columns [poly(2-hydroxyethyl methacrylate-N-methacryloyl-L-aspartic acid) (PHEMAsp)] were characterised by swelling studies, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The monolithic columns were used for IgG adsorption/desorption from aqueous solutions and human plasma. The IgG adsorption depended on the buffer type, and the maximum IgG adsorption from aqueous solution in phosphate buffer was 0.085 mg/g at pH 6.0. The monolithic columns allowed for one-step IgG purification with a negligible capacity decrease after ten adsorption-desorption cycles. (C) 2013 Elsevier B.V. All rights reserved.