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A Novel Affinity Support Material for the Separation of Immunoglobulin G from Human Plasma
Author(s) -
Garipcan Bora,
Denizli Adil
Publication year - 2002
Publication title -
macromolecular bioscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.924
H-Index - 105
eISSN - 1616-5195
pISSN - 1616-5187
DOI - 10.1002/1616-5195(20020401)2:3<135::aid-mabi135>3.0.co;2-8
Subject(s) - adsorption , aqueous solution , chemistry , (hydroxyethyl)methacrylate , swelling , human serum albumin , methacrylate , nuclear chemistry , scanning electron microscope , chromatography , polymerization , materials science , organic chemistry , polymer , composite material
2‐Methacrylamidohistidine (MAH) as a pseudospecific ligand was synthesized from methacryl chloride and histidine. Spherical beads with an average size of 50–63 μm were obtained by the radical suspension polymerization of MAH and 2‐hydroxyethyl methacrylate (HEMA) conducted in an aqueous dispersion medium. Owing to the reasonably rough character of the bead surface, P(HEMA ‐co‐ MAH) beads had a specific surface area of 17.6 m 2 ·g –1 . Synthesized MAH was characterized by NMR. P(HEMA ‐co‐ MAH) beads were characterized by swelling studies, FT‐IR spectroscopy, scanning electron microscopy (SEM) and elemental analysis. P(HEMA ‐co‐ MAH) affinity beads with a swelling ratio of 65% were used in the separation of human immunoglobulin G (HIgG) from aqueous solutions and human plasma. The maximum HIgG adsorption on the P(HEMA ‐co‐ MAH) adsorbents was observed at pH 7.4 for phosphate and at pH 6.0 for morpholinoethanesulfonic acid buffers. The HIgG adsorption onto the PHEMA adsorbents was negligible. Higher adsorption values (up to 46.5 mg·g –1 ) were obtained when the P(HEMA ‐co‐ MAH) adsorbents were used in aqueous solutions. Much higher amounts of HIgG were adsorbed from human plasma (up to 73.8 mg·g –1 ). Adsorption capacities of other blood proteins were obtained as 3.2 mg·g –1 for fibrinogen and 4.6 mg·g –1 for albumin. The total protein adsorption was determined to be 82.2 mg·g –1 . The pseudospecific affinity beads allowed one‐step separation of HIgG from human plasma. HIgG molecules could be repeatedly adsorbed and desorbed with these adsorbents without noticeable loss in their HIgG adsorption capacity.