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KMID : 1007519990080060367
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Food Science and Biotechnology
1999 Volume.8 No. 6 p.367 ~ p.371
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Metal Affinity Separation of Proteins in Aqueous PEG/Sulfate Two - Phase Systems
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Lee, Ho
Kwon, Yun Joong/Kim, Jang Hyun
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Abstract
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The feasibility of an integrated affinity adsorption-two phase extraction process was investigated using Cu(Ⅱ)-bound Sepharose particles in an aqueous PEG/sulfate two-phase system. The metal-protein coordination was not disrupted by high concentrations of phase-forming salts, thereby making it possible for this process to be performed in low-cost polymer/salt two-phase systems preferred for large scale applications. The particles which were partitioned in the interface were separated from the two-phase system, and the bound protein was recovered efficiently with imidazole treatment. By adding small amounts of imidazole during the binding process, hemoglobin with many accessible histidine residues could be eluted efficiently from metal chelate-protein complexes. This process can be applied successfully to the isolation of the desired protein from crude homogenates, with high selectivity, in a single step by the use of genetically-engineered affinity tail.
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