Advances in Slalom Chromatography for Gene Therapy Analysis

Strategic Insights -

Researchers at Waters Corporation have made significant strides in the analysis of cell and gene therapy drug substances using slalom chromatography (SC). The latest UHPLC columns and systems, combined with a better understanding of SC's separation and mass transfer mechanisms, have demonstrated its potential in analyzing plasmid topology, DNA restriction mapping, and CRISPR-Cas9 ribonucleoprotein assays. The team evaluated a novel SC column, the Waters GTxResolve 250 angstrom Slalom Column, and found it to be a high-performance, UHPLC-compatible alternative to agarose gel electrophoresis (AGE).

Key Takeaways:

  • Researchers at Waters Corporation have developed a novel slalom chromatography column, Waters GTxResolve 250 angstrom Slalom Column, for the analysis of cell and gene therapy drug substances.
  • The column demonstrated superior retention and efficiency compared to traditional agarose gel electrophoresis (AGE) in DNA separation, with improvements in accuracy (+/- 0.3%), speed (x 20), sensitivity (x 10), and peak capacity (x 2).
  • The study evaluated the retention and efficiency of the novel SC column across a wide range of flow rates (0.1-1.5 mL/min), tris-acetate-EDTA (TAE) buffer concentrations (4-400 mM, pH 8.2), and temperatures (23-80 degrees C).
  • The research demonstrated the potential of SC in analyzing plasmid topology, DNA restriction mapping, and CRISPR-Cas9 ribonucleoprotein assays, and facilitated the collection of sample fractions for secondary analyses.
  • The novel SC platform offers a high-performance, UHPLC-compatible alternative to AGE, improving ease of use, accuracy, speed, sensitivity, and peak capacity for DNA analysis in cell/gene therapy workflows.

Statistics:

  • 20-fold increase in speed compared to traditional agarose gel electrophoresis (AGE).
  • 10-fold increase in sensitivity compared to AGE.
  • 2-fold increase in peak capacity compared to AGE.
  • +/- 0.3% accuracy achieved with the novel SC column.
  • 0.1-1.5 mL/min flow rates evaluated for the novel SC column.
  • 4-400 mM, pH 8.2 TAE buffer concentrations evaluated for the novel SC column.
  • 23-80 degrees C temperatures evaluated for the novel SC column.
  • 10,000 psi column back pressure achieved with the novel SC column.

Sources:

  • NewsRx. Study Results from Waters Corporation in the Area of Gene Therapy Reported (Retention and Efficiency of a Novel Slalom Chromatography Column: an Alternative To Agarose Gel Electrophoresis for Dna Separation). Biotech Week. October 29, 2025; p 327.
  • Retention and Efficiency of a Novel Slalom Chromatography Column: an Alternative To Agarose Gel Electrophoresis for Dna Separation. Journal of Chromatography A, 2025;1761.