Abstract
AFFINITY chromatography, which uses a specific ligand coupled to a solid matrix to adsorb selectively a macromolecule, has been used extensively in the purification of soluble proteins1. Although the purification of cells or cellular membrane fragments containing surface receptors has been attempted with this procedure2–15, the approach has been less successful because of difficulties in eluting bound particulate substances from the solid matrix. Recently, we described a method called affinity partitioning16 for separating soluble proteins in aqueous polymer two-phase systems17 by adding a polymfer-ligand with a relatively high affinity for a binding site on the protein to be purified, and a solubility preference for one of the phases. Since cells and cell fragments18,19 can be partitioned and recovered from aqueous polymer two-phase systems, it seemed possible that the principle of affinity partitioning could be used in their fractionation. With the absence of a solid matrix, problems of recovery would be obviated. We describe here the use of polymer coupled to a ligand that binds to the nicotinic cholinergic receptor site to purify cholinergic receptor enriched membranes derived from electroplax of Torpedo californica.
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FLANAGAN, S., TAYLOR, P. & BARONDES, S. Affinity partitioning of acetylcholine receptor enriched membranes and their purification. Nature 254, 441–443 (1975). https://doi.org/10.1038/254441a0
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DOI: https://doi.org/10.1038/254441a0
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