Aqueous Twophase Affinity Partitioning

Affinity partitioning is an inexpensive alternative to affinity chromatography-based purification technologies, and it is also suitable for large scale application. When aqueous solutions of two high molecular weight polymers, e.g. dextran and polyethylene glycol (PEG), are mixed and then left to settle, depending on the relative molecular mass of the polymers, their concentration and temperature, two phases may be formed: an upper PEG-rich phase and a bottom dextran-rich phase. Under appropriate conditions, many proteins partition to the bottom dextran-rich phase. However, when some of the PEG of the upper phase is replaced by a suitable dye-PEG conjugate, the targeted protein may bind to the dye-PEG conjugate and selectively partition to the upper PEG-rich phase, thus, leading to some purification. An important advantage of affinity partitioning is that the technique can be exploited at the first stage of a purification process, immediately after cell disintegration, without need of centrifugation for prior debris removal. Dye-PEG conjugates have been employed in two-phase affinity systems for partial purification of numerous enzymes, especially those recognizing nucleotides.

Solar Panel Basics

Solar Panel Basics

Global warming is a huge problem which will significantly affect every country in the world. Many people all over the world are trying to do whatever they can to help combat the effects of global warming. One of the ways that people can fight global warming is to reduce their dependence on non-renewable energy sources like oil and petroleum based products.

Get My Free Ebook


Post a comment