Use of the avidin (imino)biotin system as a general approach to affinity precipitation.
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.
Freitag
Ruth
R
University of Bayreuth, Bayreuth, Germany.
Hilbrig
Frank
F
D000180:Acrylic Resins / Q000737:chemistry; D000345:Affinity Labels / Q000737:chemistry; D001360:Avidin / Q000737:chemistry*; D001710:Biotin / Q000031:analogs & derivatives* / Q000737:chemistry; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D008970:Molecular Weight; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000180:Acrylic Resins; D000345:Affinity Labels; C518163:iminobiotin; D001360:Avidin; C052970:poly-N-isopropylacrylamide; D001710:Biotin