A Novel High Resolution Silica-Based 2 µm Nonporous ODS Column
Porous ODS columns are the most popular reversed-phase columns used in today's laboratories.
Itaru Yazawa and Bryan Evans, Imtakt USA
Porous ODS columns are the most popular reversed-phase columns used in today's laboratories. Unfortunately, there are several shortcomings for porous ODS columns for polymer applications, including: poor peak shape of solutes due to wide range in pore size distribution; poor recovery of solutes due to micro-pores and meso-pores; and reduced column efficiency due to high mass transfer resistance. We have developed a 2 μm nonporous high resolution ODS column, Presto FF-C18, which can overcome these shortcomings for polymer separation.
Figure 1
Experimental and Results
All data was generated with semi-micro HPLC system equipped with UV or ELS detection. Figure 1 shows peptide separations (try ptic diges t of α-cas ein). Twice as many peaks were generated on Presto FF-C18 (2 μm nonporous) than with the 3 μm porous ODS column. Due to the nature of nonporous material (no diffusion in pores), slower flow rates can be used, which result in an increase in resolution and sensitivity. Figure 2 shows Immunoglobulin G (IgG) antibody analysis. Presto FF-C18 can provide improved peak shape and recognition between mAb (homogenous) and polyclonal antibody (heterogeneous). Figure 3 shows Hyaluronic acid analysis. Using gradient elution, Presto FF-C18 shows improved peak shape over SEC columns for polysaccharide analysis.
Figure 2
Conclusion
Presto FF-C18 shows improved resolution, peak shape, and sensitivity for polymer analysis. Presto FF-C18 can expand the separation possibilities when complimented with porous ODS columns.
Figure 3
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SEC-MALS of Antibody Therapeutics—A Robust Method for In-Depth Sample Characterization
June 1st 2022Monoclonal antibodies (mAbs) are effective therapeutics for cancers, auto-immune diseases, viral infections, and other diseases. Recent developments in antibody therapeutics aim to add more specific binding regions (bi- and multi-specificity) to increase their effectiveness and/or to downsize the molecule to the specific binding regions (for example, scFv or Fab fragment) to achieve better penetration of the tissue. As the molecule gets more complex, the possible high and low molecular weight (H/LMW) impurities become more complex, too. In order to accurately analyze the various species, more advanced detection than ultraviolet (UV) is required to characterize a mAb sample.
