Kinetic Efficiency and Fractionation Range of GTxResolve™ Premier SEC 1000 Å 3 μm Columns

Význam tématu

Size exclusion chromatography (SEC) is essential for characterizing size variants and aggregates in biopharmaceuticals, notably nucleic acid–based therapeutics such as mRNA vaccines and gene therapy vectors. Reliable SEC analysis underpins quality control of high molecular weight species and integrity assessment of DNA and RNA constructs.

Cíle a přehled studie / článku

This study evaluates the kinetic efficiency and fractionation range of the new GTxResolve™ Premier SEC 1000 Å 3 µm column, comparing it to a conventional 5 µm, 1000 Å material. DNA and RNA ladders serve as model analytes to benchmark performance across the sizing range.

Použitá metodika a instrumentace

• LC system: ACQUITY UPLC H-Class Plus Bio with binary solvent management
• Detector: Tunable UV at 260 nm and 230 nm
• Column: GTxResolve Premier SEC 1000 Å 3 µm, 150×4.6 mm; comparison with 5 µm silica 1000 Å column
• Mobile phase: 2×PBS buffer, sterile filtered
• Flow rates: 0.05–0.60 mL/min; column temp 25 °C; sample 6 °C
• Analytes: dsDNA ladders (50–1350 bp), ssRNA ladders

Hlavní výsledky a diskuse

Calibration curves for DNA and RNA showed nearly identical fractionation ranges and pore structure for both columns. However, plate height measurements demonstrated that the 3 µm column achieved 1.5–2× lower H values, translating into 1.2–1.4× higher resolution or equivalent separation in 30–50 % shorter time. Separation impedance analysis confirmed lower resistance on the 3 µm column, especially at low flow rates for small DNA and at high flow rates for larger molecules. Kinetic plots at a fixed 100 bar pressure illustrated that achieving 20 000 plates required less than half the analysis time with the 3 µm material. Example separations of DNA ladders at 0.05 mL/min showed nearly double the plate numbers and improved peak resolution across the ladder.

Přínosy a praktické využití metody

• Up to 2× faster analyses or 30–40 % higher resolution
• Reduced sample consumption due to shorter column length
• Improved reliability in quantitating high molecular weight species with minimal secondary interactions
• Scalable operating conditions for diverse nucleic acid sizes (100 nt to several kb)

Budoucí trendy a možnosti využití

As nucleic acid therapeutics diversify, the demand for robust SEC platforms will grow. The 3 µm ultra-wide pore technology can be extended to LNP aggregates, topological variants of plasmids, and emerging modalities. Further optimization of pore size and surface chemistry will support personalized mRNA vaccines and high-throughput QC workflows.

Závěr

The GTxResolve™ Premier SEC 1000 Å 3 µm column offers a significant advance in SEC performance, combining ultra-wide pores with small particle size to deliver faster, higher-resolution separations. Its superior kinetic efficiency and reduced separation impedance enable more efficient characterization of nucleic acid therapeutics compared to conventional 5 µm SEC columns.

Reference

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