Affinity chromatography in nonionic detergent solutions.

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Abstract
Anionic dye affinity chromatography is commonly unproductive in the presence of nonionic detergents used to extract particulate proteins. Using [rabbit muscle] lactate dehydrogenase (EC 1.1.1.27) as a model protein, Cibacron blue F3GA as a model dye and Triton X-100 as a model detergent, it is found that the dye is encapsulated in nonionic detergent micelles, rendering the dye incapable of ligation with the enzyme. The dye can be liberated from the micelles without altering the nonionic detergent concentration by addition of an anionic detergent, such as deoxycholate or sodium dodecyl sulfate, forming mixed anionic/nonionic micelles that displace the anionic dye. Encapsulation of the anionic detergents prevents their activity as protein denaturants. These observations were successfully translated to the dye affinity chromatography of a detergent extract of [bovine] brain particulate cyclic nucleotide phosphodiesterase.