THE USE OF ZWITTERGENT-3-14 IN THE PURIFICATION OF RECOMBINANT HUMAN INTERFERON-BETA SER(L7) (BETASERON)

A new method for purifying human interferon-beta SER(17) from E, coli-derived inclusion bodies has been developed, This procedure eliminates the need for strong denaturants, such as sodium dodecyl sulfate or chaotropes, The procedure makes use of a nondenaturing detergent and a brief incubation at pH 12 to solubilize interferon-beta Se-17 from inclusion bodies, The detergent used was Zwittergent 3-14 (nonionic and pH-insensitive), which is included in the class of sulfobetaines (RN(+)(CH3)(2)(CH2)(x)SO3-). Zwittergent 3-14 was used in combination with urea to produce a urea/Zwittergent 3-14 washed inclusion body preparation enriched in human interferon-beta Se-17 (Betaseron). Solubilization of inclusion bodies was accomplished by employing a brief (1 minute) shift to pH 12 in the presence of 2.5% Zwittergent 3-14 followed by rapid adjustment to pH 8.0. Solubilization was complete, and the solution could be rapidly adjusted to pH 8 without any observable precipitation of protein, The resultant supernatant could be successfully subjected to a number of chromatographic and analytic procedures, many of which are not compatible with strong anionic detergents, such as SDS. Betaseron was purified from Zwittergent 3-14 solubilized inclusion body lysates using both ion-exchange and size-exclusion chromatography, Purified Betaseron retained bioactivity and could be refolded by simple dialysis against a nonreducing buffer, This method represents a novel procedure for purifying Betaseron from inclusion bodies using a nondenaturing detergent and ion-exchange chromatography.,