Ion exchange chromatography (IEX) is widely used for Bio QC methods, especially for charged variant analyses. One of the critical requirements for any QC method is a reliable lot-to-lot reproducibility.
In contrast to many other IEX columns on the market, including those used as "standard", YMC's BioPro IEX columns offer excellent lot-to-lot reproducibility. On top of this comes the superior resolution. These properties make the columns the first choice in Bio QC.
Special challenges
Sensitive substances like oligonucleotides tend to interact with column hardware, even with quite inert IEX standard hardware made of PEEK. In addition, diverse peptides and proteins are prone to interaction with the column body, too.
Adsorption of the analytes on the surfaces leads to peak tailing, carryover, and reduced recovery. In the past, time and solvent-intensive preconditioning was the only measure. Fortunately, suitable bioinert column hardware makes it much easier nowadays.
Bioinert column hardware as a solution
YMC meets this special challenge with the recently launched YMC Accura columns. They feature a bioinert surface coating of the column body and frits.
The robust bioinert coating used on YMC Accura hardware is 130 to 320-fold thicker in comparison to similar hardware concepts.
Permanent inertness against sensitive substances is ensured. The greatest achievement is of course, that YMC Accura columns can even be used for the analysis of sensitive substances without pre-conditioning.
Figure 1: YMC Accura BioPro IEX columns feature a bioinert coating of the column body and frits. © YMC Europe
Ideal stationary phases
The non-porous hydrophilic polymer beads of the BioPro IEX stationary phases enable exceptionally high efficiencies and high throughput analyses. Even small sample variations can be easily separated and detected. The superior lot-to-lot reproducibility guarantees highly reliable results.
BioPro IEX SF with its strong cation exchanger functionality is particularly ideal for analyzing charge variants of monoclonal antibodies (mAbs) or other proteins.
On the other hand, the strong anion exchanger BioPro IEX QF is the first choice for high-resolution analysis of oligonucleotides of different lengths as well as mAbs with lower pI.
Figure 2: YMC Accura BioPro IEX columns provide high recovery and good peak shapes from the first injection making them ideal for the analysis of oligonucleotides (here: 21mer RNA) with reproducible results. © YMC Europe
Benefits of using YMC Accura BioPro IEX columns
The combination of the bioinert coated column hardware and the high-efficiency stationary phases of YMC Accura BioPro IEX columns provide excellent peak shapes for sensitive compounds. They ensure high recovery from the very first injection and at the same time prevent carry-over.
This advantage stands out especially when analyzing oligonucleotides compared to conventional PEEK hardware (Figure 2). YMC Accura BioPro IEX columns are not only very inert, but are also available with smaller column IDs, making them ideal for use in native IEX MS analysis.
For example, Figure 3 shows the high-sensitivity analyses of a monoclonal antibody using a YMC Accura BioPro IEX SF column with a 3 µm particle size and column dimensions of 100 x 2.1 mm ID.
Figure 3: Smaller column IDs that allow lower flow rates and volatile mobile phases are necessities for coupling to MS. Such high-sensitivity analyses of mAbs (here: trastuzumab) are ideally performed using YMC Accura BioPro IEX columns. © YMC Europe
This article was originally published in Analytical Science.
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