Biopharmaceutical Analysis
From mAbs to Oligos, We Have You Covered
Phenomenex is committed to the development of novel chromatographic solutions that advance biopharmaceutical development. This rapidly-growing industry necessitates fit for purpose applications and innovative technologies to meet the expanding analytical challenges of the field.
Phenomenex columns and sample preparation solutions are designed to tackle the multi-faceted analytical needs of large molecule analysis:
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Check out our most commonly asked Biopharmaceutical Analysis Questions.
What is the loading capacity of bioZen Peptide and Intact columns?The bioZen Peptide columns have similar loading capacities as reversed phase HPLC/UHPLC columns. A 5-20 μg of digest or peptide mixture on a 4.6 mm ID column will provide good sensitivity especially for LC-MS peptide separations. Up to 50 μg of digest can be loaded without increasing peak width too severely. For 2.1 mm ID columns, loading should be scaled accordingly. For bioZen Intact columns, overloading can drastically affect peak shape due to their lower surface area and must be determined experimentally for optimal results. For 4.6 mm ID’s, 5 μg is a good starting point. For 2.1 mm ID’s, 1 μg is a recommended starting point. Increasing in load may increase peak tailing and peak width significantly. |
What are some mobile phase considerations for Size Exclusion Chromatography/GFC?The most common application for SEC is aggregate analysis, which requires “native” conditions (i.e. physiological conditions). These conditions require a buffered solution at around pH 7 with a moderate amount of salt. A good starting point for mobile phase is a 100 mM Phosphate Buffer, pH 6.8. However, there are some instances where it is appropriate to optimize the mobile phase to improve chromatography. If you suspect that the protein is hydrophobic, a small amount of organic solvent (e.g. methanol, acetonitrile, or isopropanol) can be added generally anywhere between 5-10 %. If you suspect that the protein may have strong ionic characteristics (e.g. basic protein or protein with a high isoelectric point), the ionic strength of the eluent can be increased to improve chromatography. A 2x phosphate buffered saline (i.e. 50 mM phosphate, 300 mM sodium chloride, pH 6.8) is recommended to prevent secondary interactions. Note that increasing the salt will also increase hydrophobic interactions. Further, varying the salt may improve the separation of one critical pair, but result in a loss of resolution of another. |
