Abstract

Sephacryl is a gel filtration medium composed of allyl dextran and N,N'-methylene bisacrylamide. Sephacryl S-200 SF has a fractionation range similar to that of Sephadex G-150. Sephacryl S-300 SF has a fractionation range more similar to Sephadex G-200 or Sepharose 6B, with an exclusion limit for globular proteins of about 10⁶ daltons. In typical applications, excellent resolution of protein mixtures was obtained at flow rates up to 30 cm/hr. Good correlation was established between elution volumes of proteins and their molecular weights. The gels contain few ionic groups, as determined by titration. Studies using Sephacryl S-200 at different pH values indicate that, above pH 3, protein does not interact with Sephacryl S-200 at moderate ionic strengths. However, below pH 3, protein binds strongly to the gel through hydrophobic interactions and/or hydrogen bonding. Most separations using Sephadex G-150 or Sephadex G-200 could be duplicated using Sephacryl S-200 or S-300 at much higher flow rates.