Novel metal-chelate affinity adsorbent for purification of immunoglobulin-G from human plasma

Metal-chelating ligand and/or comonomer 2-methacrylolyamidohistidine (MAH) was synthesized by using methacryloyl chloride and l-histidine methyl ester. MAH was characterized by NMR and FTIR. Spherical beads with an average diameter of 75–125 μm were produced by suspension polymerization of methylmet...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2003-09, Vol.795 (1), p.93-103
Main Authors: Denizli, Adil, Alkan, Murat, Garipcan, Bora, Özkara, Serpil, Pişkin, Erhan
Format: Article
Language:English
Subjects:
Adsorption
Analytical biochemistry: general aspects, technics, instrumentation
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chelating Agents - chemistry
Chromatography, Affinity - methods
Fundamental and applied biological sciences. Psychology
Humans
Hydrogen-Ion Concentration
Immunoglobulin G
Immunoglobulin G - blood
Immunoglobulin G - isolation & purification
Magnetic Resonance Spectroscopy
Metals - chemistry
Microscopy, Electron, Scanning
Spectroscopy, Fourier Transform Infrared
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Summary:Metal-chelating ligand and/or comonomer 2-methacrylolyamidohistidine (MAH) was synthesized by using methacryloyl chloride and l-histidine methyl ester. MAH was characterized by NMR and FTIR. Spherical beads with an average diameter of 75–125 μm were produced by suspension polymerization of methylmethacrylate (MMA) and MAH carried out in an aqueous dispersion medium. Poly(MMA-MAH) beads had a specific surface area of 37.5 m 2/g. Poly(MMA-MAH) beads were characterized by water uptake studies, FTIR, SEM and elemental analysis. Elemental analysis of MAH for nitrogen was estimated as 34.7 μmol/g of polymer. Then, Cu 2+ ions were chelated on the beads. Cu 2+-chelated beads with a swelling ratio of 38% were used in the adsorption of human-immunoglobulin G (HIgG) from both aqueous solutions and human plasma. The maximum adsorption capacities of the Cu 2+-chelated beads were found to be 12.2 mg/g at pH 6.5 in phosphate buffer and 15.7 mg/g at pH 7.0 in MOPS. Higher adsorption value was obtained from human plasma (up to 54.3 mg/g) with a purity of 90.7%. The metal-chelate affinity beads allowed one-step separation of HIgG from human plasma. The adsorption–desorption cycle was repeated 10 times using the same beads without noticeable loss in their HIgG adsorption capacity.
ISSN:1570-0232
1873-376X
DOI:10.1016/S1570-0232(03)00550-6