Nylon-6 capillary-channeled polymer (C-CP) fibers as a hydrophobic interaction chromatography stationary phase for the separation of proteins

• Published in: Analytical and bioanalytical chemistry 2009-01, Vol.393 (1), p.273-281
• Main Authors: Stanelle, Rayman D, Marcus, R. Kenneth
• Format: Article
• Language: English
• Subjects: Analytical Chemistry; Biochemistry; Capillary-channeled polymer; Caprolactam - analogs & derivatives; Caprolactam - chemistry; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatographic methods and physical methods associated with chromatography; Chromatography; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; Exact sciences and technology; Fibers; Flow rate; Food Science; Hydrophobic and Hydrophilic Interactions; Hydrophobic interaction chromatography; Laboratory Medicine; Mathematical analysis; Molecular Structure; Monitoring/Environmental Analysis; Muramidase - analysis; Muramidase - isolation & purification; Nylons; Optimization; Original Paper; Other chromatographic methods; Permeability; Polymers - chemistry; Protein; Proteins; Ribonuclease, Pancreatic - analysis; Ribonuclease, Pancreatic - isolation & purification; Separation; Surface Properties; Time Factors; Transferrin - analysis; Transferrin - isolation & purification
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-008-2457-2

Nylon-6 capillary-channeled polymer (C-CP) fibers are used as the stationary phase for the hydrophobic interaction chromatography (HIC) separation of a synthetic protein mixture composed of ribonuclease A, lysozyme, and holotransferrin. Nylon is a useful polymer phase for HIC as it has an alkyl backbone, while the amide functionality is hydrophilic (in fact ionic) in nature. The combination of a nonporous polymer surface of the fiber phases and high column permeability yields very efficient mass transfer characteristics, as exhibited by narrowing of peak widths with increases in mobile phase linear velocity. Retention factors and resolution were evaluated at flow rates ranging from 0.5 to 9 mL/min (linear velocities of ca. 2 to 15 mm/s) and at gradient slopes between 3.3 and 20 %B/min. Optimum resolution was achieved by employing fast flow rates (9 mL/min) and slow gradients (3 %B/min), also resulting in the highest peak capacities.