A Novel Magnetic Affinity Support for Protein Adsorption and Purification

A novel magnetic support was prepared by an oxidization‐precipitation method with poly(vinyl alcohol) (PVA) as the entrapment material. Transmission electron microscopy indicated that the magnetic particles had a core‐shell structure, containing many nanometer‐sized magnetic cores stabilized by the...

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Bibliographic Details
Published in:Biotechnology progress 2001, Vol.17 (1), p.134-139
Main Authors: Tong, Xiao-Dong, Xue, Bo, Sun, Yan
Format: Article
Language:English
Subjects:
Adsorption
Alcohol Dehydrogenase - chemistry
Alcohol Dehydrogenase - isolation & purification
Biological and medical sciences
Biotechnology
Chromatography, Affinity - methods
Fundamental and applied biological sciences. Psychology
Kinetics
Magnetics
magnetism
Methods. Procedures. Technologies
Muramidase - chemistry
Muramidase - isolation & purification
Osmolar Concentration
Others
protein purification
Various methods and equipments
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Summary:A novel magnetic support was prepared by an oxidization‐precipitation method with poly(vinyl alcohol) (PVA) as the entrapment material. Transmission electron microscopy indicated that the magnetic particles had a core‐shell structure, containing many nanometer‐sized magnetic cores stabilized by the cross‐linked PVA. The particles showed a high magnetic responsiveness in magnetic field, and no aggregation of the particles was observed after the particles had been treated in the magnetic field. These facts indicated that the particles were superparamagnetic. Cibacron blue 3GA (CB) was coupled to the particles to prepare a magnetic affinity support (MAS) for protein adsorption. Lysozyme was used as a model protein to test the adsorption properties of the MAS. The adsorption equilibrium of lysozyme to the MAS was described by the Langmuir‐type isotherm. The capacity for lysozyme adsorption was more than 70 mg/g MAS (wet weight) at a relatively low CB coupling density (3−5 μmol/g). In addition, 1.0 M NaCl solution could be used to dissociate the adsorbed lysozyme. Finally, the MAS was recycled for the purification of alcohol dehydrogenase (ADH) from clarified yeast homogenates. Under proper conditions, the magnetic separation yielded over 5‐fold purification of the enzyme with 60% recovery of the enzyme activity.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp000134g