GTxResolve SEC Columns

Platform multi-attribute quantitation with SEC-MALS

Adeno-associated virus (AAV) and other viral vectors are promising modalities for treatment of rare diseases. To ensure the safety and efficacy of these drugs, the drug content and level of impurities need to be measured. SEC is a promising separation technique for assessing the amount of the sample’s different components.

This application demonstrates the assessment of degradation pathways for an AAV using a novel and robust surface chemistry particle. The lower secondary interactions and high efficiency of the GTxResolve SEC 1000 Å Column and GTxResolve Premier BEH SEC 450 Å Column facilitate and elucidate new drug design insights and can therefore be used as a suitable tool as starting point for platform methods.

Integrity analysis for small to larger nucleic acids

Integrity studies involving large nucleic acids, such as mRNA, can benefit from a quick size exclusion analysis. SEC with a GTxResolve Premier SEC 1000 Å Column makes it possible to achieve high efficiency separations and quickly assess the integrity and stability of larger nucleic acids. In this application, the GTxResolve Premier SEC 1000 Column is paired with MALS detection, which demonstrates the utility of wide pore SEC for biophysical attribute characterization.

LNP heterogeneity with wide pore technology

Lipid nanoparticles (LNPs) are critical carriers for mRNA delivery in vaccines and gene therapies. This application uses the Waters GTxResolve 2000 Å SEC Column with MaxPeak Premier 3 µm particles to compare the heterogeneity of LNP formulations from two different manufacturers. The chromatogram reveals distinct elution profiles: Manufacturer 1 shows a narrow, symmetrical peak, indicating a relatively homogeneous LNP population. Manufacturer 2 presents a broader elution profile with additional shoulders and secondary peaks, suggesting a more heterogeneous formulation with larger particle sizes. The ability to profile such differences highlights the column’s sensitivity and resolving power, supporting its use in both development and quality control of LNP-based therapeutics.

Resolving plasmids and vectors

Characterizing nucleic acid therapeutics requires precise tools capable of resolving closely related DNA species. This is especially important in applications where size and topology directly influence potency, transfection efficiency, and regulatory compliance. Here, we demonstrate the high-resolution separation of two structurally similar circular DNA species. These molecules are difficult to separate using conventional SEC columns and, in this example, the Waters GTxResolve 2000 Å SEC Column achieves baseline resolution, especially between pBR322 and its related species as highlighted in the circled region, and clearly distinguishing between the two DNA constructs.

Size characterization of an intact lentiviral vector (LVV)

Lentiviruses (LVs) are a type of retrovirus widely used as vectors in gene therapy, cell therapy (like CAR-T), and vaccine development. What makes them particularly valuable is their ability to stably integrate genetic material into both dividing and none-dividing cells, allowing for long-term expression of therapeutic genes. Characterization of LV is crucial to ensure its potency, purity, and safety.  It provides key insights into particle integrity, concentration, and aggregation, critical for developing robust, scalable, and regulatory-compliant gene therapy success.  Here, we demonstrate separation of a downstream processed LV sample (Rt 12min) from other fragmented proteins (Rt 14-15.5) generated during processing using Waters GTxResolve 2000Å SEC Column, MaxPeak 3µm.

Drug stability and integrity study for mRNA-LNP based vaccines

Scientists must monitor a drug’s integrity and stability due to the implications on safety and efficacy. Reproducibility, purity, specificity, and linear range are some of the critical quality attributes considered for mRNA-based vaccines. Size exclusion chromatography is a well-suited technique for characterization of larger mRNA and LNP aggregates in nucleic-acid-based vaccines. The consistency of lipid nanoparticle (LNP) content and linear response of two LNP formulations (X1 & X2) from two different manufacturers using GTxResolve Premier SEC 1000 Å was investigated. In the example applications, the linear response is measured by plotting the peak area against the amount of LNP injected onto the GTxResolve Premier SEC 1000 Column.