Liquid-liquid extraction of a recombinant protein with a reverse micelle phase.

Recombinant cytochrome b5 was extracted to a reversed micelle phase of a cationic surfactant (hexadecyltrimethylammonium bromide, CTAB) in cyclohexane/decanol and back-extracted to a fresh aqueous phase. Depending on the experimental conditions, i.e., temperature, pH, and ionic strength, the extraction is controlled by either hydrophobic (pH close to the pI and high ionic strength) or electrostatic (pH far from the pI and low ionic strength) interactions. The protein interacts strongly with the surfactant aggregate once it is extracted to the organic phase, and it is very difficult to back-extract it to a new aqueous phase.