Dynamic evaluation of a trilobal capillary‐channeled polymer fiber shape for reversed phase protein separations and comparison to the eight‐channeled form

Abstract

A new, trilobal-shaped capillary-channeled polymer fiber is under development to address the issues of poor A-term performance of the previous eight-channeled form. The trilobal geometry should provide better packing homogeneity due to the fewer potential orientations of the symmetric fiber geometry. Comparisons of separation efficiency and peak shape were made between the two fiber shapes through several dynamic parameters. Column hydrodynamics were investigated with two marker compounds, uracil and bovine serum albumin, with van Deemter plots of those two compounds revealing differences in the packing qualities between the different fiber shapes. Parametric fitting to the van Deemter, Knox, and Giddings equations provides insights into the column physical structures. Separation quality for both shapes was evaluated across differences in fiber packing density, gradient rate, and mobile phase linear velocity for the reversed phase separation of a four protein mixture, containing ribonuclease A, cytochrome c, lysozyme, and myoglobin. The results of this study lay the ground work for future efforts in the use of trilobal fibers for the separation of biomacromolecules.

MICA-G129R: A bifunctional fusion protein increases PRLR-positive breast cancer cell death in co-culture with natural killer cells

“…Using the nickel resin chromatography, fusion protein MICA-G129R was successfully purified from the conditioned media. HPLC analysis of the purified MICA-G129R using a C-CP fiber column [ 34 , 35 ] revealed that the peak representing MICA-G129R (indicated with the white arrow in Fig 4A ) dramatically increased to 70.0% of the total area under the curve, meaning that the purity of MICA-G129R in the purified solution was 70.0%. The other peaks dampened or disappeared after the purification, which reflected that most of other proteins in the conditioned media were removed.…”

“…Polypropylene trilobal capillary-channeled polymer (C‐CP) fibers were packed as the HPLC column as previously described [ 34 ]. The separation procedures were carried out following a method built previously [ 35 ].…”

Comparison of the separation of proteins of wide‐ranging molecular weight via trilobal polypropylene capillary‐channeled polymer fiber, commercial superficiously porous, and commercial size exclusion columns

“…The PPY C-CP fiber column manufacture has been described in previous communications [29,38]. Briefly, eight rotations of PPY fibers (∼560 fibers) were heat shrunk using boiling water and then cleaned sequentially using room temperature Milli-Q water, ACN, and Milli-Q water to remove any latent anti-static spin-coating.…”

“…Likewise, the absence of appreciable phase porosity versus the size of proteins results in separations with virtually no van Deemter Cterm effects [29,37], a critical point in terms of separations of proteins of increasing molecular weight. Recently, a Y-shaped trilobal, polypropylene fiber structure (referred to as PPY) was described and its performance compared to the typical eight-channeled shape for reversed-phase protein separations [23,38]. Ultimately, with more uniform packing (lower van Deemter A-term), the PPY columns yield elution peaks with greater symmetry and higher separation efficiency (resolution) than the previous eight-channel fiber geometry.…”

Comparative analysis of trilobal capillary‐channeled polymer fiber columns with superficially porous and monolithic phases toward reversed‐phase protein separations

“…A number of detailed studies using C-CP fiber phase separations of proteins point to the van Deemter A-term being the primary limiting aspect, with C-term broadening being virtually non-existent at linear velocities of up to 100 mm/s [60,62,63]. The lack of C-term broadening is a direct result of the non-porous nature of the fiber surfaces with respect to the size of proteins as well as enhanced mobile phase mass transfer at relatively high linear velocities [60,61,64].…”