Immobilized metal-ion affinity chromatography: Imidazole proton pump and chromatographic sequelae. II. Chromatographic sequelae

✍ Scribed by Eugene Sulkowski

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 469 KB
• Volume: 9
• Category: Article
• ISSN: 0952-3499
• DOI: 10.1002/(sici)1099-1352(199634/12)9:5/6<494::aid-jmr290>3.0.co;2-f

✦ Synopsis

Isocratic elution with imidazole of some protein models from a chelating gel, CSFF-IDA-M(II), resulted in their desorption owing to the low pH (644) of the mobile phase rather than to the imidazole itself (idazole-generated fall in pH; proton pump). Gradient elution with imidazole was best accomplished when the chelating gel was initially converted into its imidazole complex, CSFT-IDA-MOI)Im. The exploitation of the imidazole-generated proton pump in the IMAC of proteins may enhance the versatility of this type of chromatography.

Keywordr: immobilized metal-ion affinity chromatography; imidazole; pH elution; iminodiacetate-metal chelate proton pump NaCl), pH 4.0, and the columns were subsequently equilibrated with 10-20 m~ phosphate buffer (PB), containing 1 M NaCl, pH 7.0. Protein samples, containing 5-10 mg