summary
Biospecific interactions are used in many capturing and bioseparation steps. A typical situation is the coupling of a biospecific ligand to a chromatographic stationary phase for affinity chromatography. This approach has two possible drawbacks. The first is that a chromatographic column may be awkward to use in experimental setups; the second is related to the need to develop a dedicated coupling chemistry for any given affinity ligand. In affinity precipitation, the biospecific affinity ligand is instead linked to a stimuli-responsive molecule to yield a so-called affinity macroligand (AML). Upon stimulation, such molecules show abrupt yet reversible precipitation from aqueous solution. Capture by affinity precipitation just requires the addition of the stimuli-responsive AML to the raw target solution followed by selective precipitation of the formed affinity complex via the application of the stimulus. The need for the synthesis of a dedicated AML may be circumvented by the use of an avidin-activated stimuli-responsive precursor, to which any biotinylated affinity ligand can be securely linked via the well-known strong avidin–biotin interaction.
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Acknowledgments
We would like to thank the numerous students and collaborators who helped to develop and test out the experimental procedures, in particular Matteo Costioli, Arnaud Deponds, Frédéric Garret-Flaudy, Marilia Panayiotou, and Gisela Stocker. Our work in this area has for many years been supported by the German Science Foundation (DFG), the Swiss National Science Foundation (SNF), and the Commission for Innovation and Technology (CTI), Switzerland.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Freitag, R., Hilbrig, F. (2008). Use of the Avidin (Imino)Biotin System as a General Approach to Affinity Precipitation. In: McMahon, R.J. (eds) Avidin-Biotin Interactions. Methods In Molecular Biology™, vol 418. Humana Press. https://doi.org/10.1007/978-1-59745-579-4_4
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DOI: https://doi.org/10.1007/978-1-59745-579-4_4
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