Overload Effects in Reversed Phase Protein Separations using Capillary-Channeled Polymer Fiber Columns

Biotechnology Progress
Lei Wang, R Kenneth Marcus

Abstract

In preparative chromatography, overloading effects are a fact of life. However, peak distortion is problematic in analytical scale chromatography, especially in the case of bio-macromolecule separations. Capillary-channeled polymer fibers have been employed for fast protein separations in reversed phase, ion exchange, and hydrophobic interaction liquid chromatography. Although the primary advantage of the phase is operation at high linear velocities (~100 mm s-1 ) without van Deemter C-term limitations, the limited specific surface area (<5 m2 g-1 ) suggests that a thorough understanding of overloading effects is needed. We evaluate important factors (injected mass and volume) affecting overload in terms of peak height, width and shape (asymmetry). Overload conditions readily apparent as the peak shape changes from a Gaussian distribution to a left-triangle with slight tailing; more-or-less classical overload behavior. Three methods were used to compute column efficiency in terms of plate counts (N), including the area height, half-height, and the Dorsey-Foley approaches. The half-height method is best-suited for describing column overload, accounting for peak distortion and achieving high levels of consistency. The limiting pl...Continue Reading

References

Dec 10, 1997·Journal of Chromatography. B, Biomedical Sciences and Applications·W C Lee
Aug 1, 1995·Industrial & Engineering Chemistry Research·A Velayudhan, M R Ladisch
Apr 6, 2002·Biotechnology Progress·Chenghong LiMichael R Ladisch
Oct 31, 2003·Journal of Chromatography. a·Fabrice GrittiGeorges Guiochon
Jul 29, 2004·Biotechnology and Bioengineering·Sanchayita GhoseBrian Hubbard
Oct 1, 2005·Journal of Chromatography. a·Rayman D StanelleR Kenneth Marcus
Nov 12, 2005·Analytical and Bioanalytical Chemistry·Rayman D StanelleR Kenneth Marcus
Oct 13, 2006·Journal of Chromatography. a·R Andrew ShallikerGeorges Guiochon
Oct 5, 2010·Journal of Chromatography. a·Lena EdströmTorgny Fornstedt
Jun 28, 2012·Analytical and Bioanalytical Chemistry·K Manoj Randunu, R Kenneth Marcus
Oct 15, 2013·Biotechnology and Bioengineering·Ronald W MaurerChristopher J Roberts
Jan 1, 2014·Journal of Separation Science·Abby J Schadock-Hewitt, R Kenneth Marcus
Feb 27, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hung K TrangR Kenneth Marcus
Aug 25, 2016·Analytical and Bioanalytical Chemistry·Michael Ladisch, Leyu Zhang

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