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Affinity separation of enzymes from mixtures containing suspended solids
Authors:Bernard L Hirschbein  George M Whitesides
Institution:(1) Department of Chemistry, Massachusetts Institute of Technology, 02139 Cambridge, Massachusetts
Abstract:Agarose beads containing immobilized enzymes or affinity ligands have been made magnetically responsive by adsorbing freshly precipitated magnetite on their surface. These beads are used for affinity adsorption of proteins from complex mixtures containing suspended solids. The magnetically responsive beads and the unwanted (diamagnetic) solids are then separated by magnetic filtration. This magnetic adsorption scheme for direct affinity separation of enzymes from mixtures containing suspended solids is compared with a similar, but nonmagnetic, scheme in which the affinity matrix is supported on fiberglass cloth. The enzyme is allowed to adsorb in this matrix, and the matrix is simply removed physically from the suspension to achieve separation from the unwanted solids. The two methods seem comparable in their ability to separate a desired enzymatic activity. The magnetic methods are technically the more complex of the two, but are significantly the more rapid. The efficiency of separation of diamagnetic and ferrimagnetic solids in these biological systems by high gradient magnetic filtration is good.
Keywords:Affinity separation  of enzymes via magnetic techniques  separation  of enzymes by magnetic affinity techniques  enzymes  separation by magnetic affinity techniques  solids  affinity separation of enzymes from mixtures of suspended  magnetic separation  of enzymes from suspended solids
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