Fast protein separation by reversed-phase high-performance liquid chromatography on octadecylsilyl-bonded nonporous silica gel: II. Improvement in recovery of hydrophobic proteins
✍ Scribed by Hiroko Itoh; Noriyuki Nimura; Toshio Kinoshita; Norikazu Nagae; Mitsugu Nomura
- Publisher
- Elsevier Science
- Year
- 1991
- Tongue
- English
- Weight
- 388 KB
- Volume
- 199
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Recovery of hydrophobic proteins from an RP-HPLC column was improved using a fast-separation RP-HPLC system operated at room temperature. Hydrophobic proteins such as ovalbumin could be adequately eluted from a nonporous octadecylsilyl (C18) spherical silica gel with a particle diameter of 20 microns using steep gradient elution with a 0.1% aqueous trifluoroacetic acid-acetonitrile system at a constant flow rate of 4 ml/min. Recoveries improved under fast separation since the protein sample suffered only a slight amount of irreversible denaturation on the hydrophobic surface of the stationary phase. The fast-separation system was also applied to the separation of larger proteins such as apo-ferritin (443 kDa) and thyroglobulin (669 kDa) as well as egg white proteins.