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Comparison of analytical protein separation characteristics for three amine-based capillary-channeled polymer (C-CP) stationary phases

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Abstract

Capillary-channeled polymer (C-CP) fiber stationary phases are finding utility in the realms of protein analytics as well as downstream processing. We have recently described the modification of poly(ethylene terephthalate) (PET) C-CP fibers to affect amine-rich phases for the weak anion-exchange (WAX) separation of proteins. Polyethylenimine (PEI) is covalently coupled to the PET surface, with subsequent cross-linking imparted by treatment with 1,4-butanediol diglycidyl ether (BUDGE). These modifications yield vastly improved dynamic binding capacities over the unmodified fibers. We have also previously employed native (unmodified) nylon 6 C-CP fibers as weak anion/cation-exchange (mixed-mode) and hydrophobic interaction chromatography (HIC) phases for protein separations. Polyamide, nylon 6, consists of amide groups along the polymer backbone, with primary amines and carboxylic acid end groups. The analytical separation characteristics of these three amine-based C-CP fiber phases are compared here. Each of the C-CP fiber columns in this study was shown to be able to separate a bovine serum albumin/hemoglobin/lysozyme mixture at high mobile phase linear velocity (∼70 mm s−1) but with different elution characteristics. These differences reflect the types of protein-surface interactions that are occurring, based on the active group composition of the fiber surfaces. This study provides important fundamental understanding for the development of surface-modified C-CP fiber columns for protein separation.

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Acknowledgments

This material is based upon the work supported by the National Science Foundation Division of Chemistry under grant CHE-1307078.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Chemistry, Biosystems Research Complex, Clemson University, Clemson, SC, 29634, USA

    Liuwei Jiang & R. Kenneth Marcus

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  1. Liuwei Jiang
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  2. R. Kenneth Marcus
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Correspondence to R. Kenneth Marcus.

Additional information

Published in the topical collection Fiber-based Platforms for Bioanalytics with guest editors Antje J. Baeumner and R. Kenneth Marcus.

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Jiang, L., Marcus, R.K. Comparison of analytical protein separation characteristics for three amine-based capillary-channeled polymer (C-CP) stationary phases. Anal Bioanal Chem 408, 1373–1383 (2016). https://doi.org/10.1007/s00216-015-9000-z

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  • DOI: https://doi.org/10.1007/s00216-015-9000-z

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