title: Aspartic acid incorporated monolithic columns for affinity glycoprotein purification.
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 × 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.
authors.affiliation: Hacettepe University, Department of Chemistry, Biochemistry Division, Ankara, Turkey.
authors.forename: Canan
authors.initials: C
authors.lastname: Armutcu
authors.affiliation: Hacettepe University, Department of Chemistry, Biochemistry Division, Ankara, Turkey.
authors.forename: Nilay
authors.initials: N
authors.lastname: Bereli
authors.affiliation: Hacettepe University, Department of Chemistry, Biochemistry Division, Ankara, Turkey.
authors.forename: Engin
authors.initials: E
authors.lastname: Bayram
authors.affiliation: Hacettepe University, Department of Chemistry, Biochemistry Division, Ankara, Turkey.
authors.forename: Lokman
authors.initials: L
authors.lastname: Uzun
authors.affiliation: Anadolu University, Department of Chemistry, Biochemistry Division, Eskişehir, Turkey.
authors.forename: Rıdvan
authors.initials: R
authors.lastname: Say
authors.affiliation: Hacettepe University, Department of Chemistry, Biochemistry Division, Ankara, Turkey. Electronic address: denizli@hacettepe.edu.tr.
authors.forename: Adil
authors.initials: A
authors.lastname: Denizli
mesh_terms: D000327:Adsorption; D001224:Aspartic Acid / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000302:isolation & purification; D008024:Ligands; D008689:Methacrylates / Q000737:chemistry*; D008855:Microscopy, Electron, Scanning; D009994:Osmolar Concentration; D012212:Rheology; D012709:Serum Albumin / Q000302:isolation & purification; D012996:Solutions; D013696:Temperature
chemical_list: D006023:Glycoproteins; D007074:Immunoglobulin G; D008024:Ligands; D008689:Methacrylates; D012709:Serum Albumin; D012996:Solutions; D001224:Aspartic Acid; C005044:hydroxyethyl methacrylate
keywords: High performance liquid chromatography; IgG separation; Monolithic columns; Pseudospecific ligands