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 ch aotropes, The procedure makes use of a nondenaturing detergent and a b rief incubation at pH 12 to solubilize interferon-beta Se-17 from incl usion 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 ur ea to produce a urea/Zwittergent 3-14 washed inclusion body preparatio n enriched in human interferon-beta Se-17 (Betaseron). Solubilization of inclusion bodies was accomplished by employing a brief (1 minute) s hift to pH 12 in the presence of 2.5% Zwittergent 3-14 followed by rap id adjustment to pH 8.0. Solubilization was complete, and the solution could be rapidly adjusted to pH 8 without any observable precipitatio n of protein, The resultant supernatant could be successfully subjecte d to a number of chromatographic and analytic procedures, many of whic h are not compatible with strong anionic detergents, such as SDS. Beta seron was purified from Zwittergent 3-14 solubilized inclusion body ly sates using both ion-exchange and size-exclusion chromatography, Purif ied Betaseron retained bioactivity and could be refolded by simple dia lysis against a nonreducing buffer, This method represents a novel pro cedure for purifying Betaseron from inclusion bodies using a nondenatu ring detergent and ion-exchange chromatography.,