|Articles|November 6, 2020

Size Exclusion Chromatography Method Development of NIST mAb Using an Agilent AdvanceBio SEC 200 Å 1.9 µm Column

Learn how to perform method optimization using a bio-inert quaternary LC instrument to screen multiple mobile-phase compositions for aggregate analysis of NIST mAb.


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Figure 2. Split injection used to measure triplicate test of MTBE, hexane, o-xylene, and 1-methylnaphthalene comparing different solvents (methanol, methylene chloride) and column insertion distances starting with 0.5 mm, 5.0 mm, 10.0 mm, 15 mm, and 20.0 mm. Methanol had the best performance at the 5.0 mm insertion distance and methylene chloride looked slightly better at the 0.5 mm. We would still recommend not going below the 5.0 mm manufacturer recommended insertion distance.
Revisiting insertion distance for methanol and methylene chloride in GC analysis and confirming that insertion depth strongly affects response and reproducibility.
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