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

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 back...

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Bibliographic Details
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
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Summary: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.
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-008-2457-2