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Thermo Fisher Scientific consumables for chromatography
Technological innovation improves quality and efficiency | Conducting polymer analysis of proteins, mRNA, and AAV under a mobile phase condition
Date: 2025-05-27Read: 2
SurePac Bio 550 SEC MDI Innovation Technology
Monodisperse particles:
Uniform 3 µ m monodisperse silica gel particle technology, excellent pore size distribution, reduces the influence of diffusion effects in chromatographic separation.
Inert technology:
Advanced filler encapsulation technology shields ion exchange sites, minimizes non-specific binding, and utilizes biologically inert hardware. It is compatible with detectors such as MS, CAD, and MALS while reducing non-specific binding.
High Reproducibility
Accurate control of the packing of chromatographic columns and higher quality control release standards have achieved excellent inter batch consistency while ensuring high column efficiency.
High throughput
The 4.6mm X 150mm column is compatible with both HPLC and UHPLC systems, and can complete the analysis of a sample in 7 minutes.
Technological Innovation - Comparison of Protein Analysis Results under Different Mobile Phase Conditions
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Advanced packing encapsulation technology shields ion exchange sites and minimizes non-specific binding. The biologically inert stainless steel SurePac Bio 550 SEC MDI chromatographic column reduces non-specific binding between proteins and the column, as shown in Figure (1). The effect of changing salt concentration on the separation of 500kDa proteins under pH 6.8 conditions is studied. High concentrations of sodium chloride can slightly improve the separation between polymer and monomer, and the difference in separation between polymer and main peak is not significant when the buffer concentration is between 20-50 mmol/L. As shown in Figure (2), at 50 mmol/L NaH2PO4+In a 300 mmol/L NaCl (pH 6.8) system, the effect of adding 0-15% acetonitrile at different ratios on protein sample separation was compared. In this experiment, it was found that the chromatographic column had little hydrophobic adsorption with the sample. In the system without acetonitrile, the separation degree between the polymer and the main peak, as well as the peak to valley ratio of the polymer, were optimal. The experimental results indicate that the ion adsorption and hydrophobic adsorption between the SurePac Bio550 SEC MDi chromatographic column and the protein are very low, and the polymer and protein can be separated relatively well in a system without acetonitrile.
As shown in Figures (2), (3), (4), and (5), the SurePac Bio 550 SEC MDi column achieved good separation of 500 kDa protein, mRNA, and AAV9 monomers and oligomers within 15 minutes. The ion adsorption and hydrophobic adsorption between the sample and the chromatographic column are relatively low. In the same chromatographic mobile phase system, polymer and monomer analysis of three types of samples can be completed simultaneously. Under the biocompatible Vanquish Flex liquid chromatography system, the chromatographic column separated AAV9 with different titers, and no residue was observed under both UV and fluorescence detectors.
References:
https://www.thermofisher.com/order/catalog/product/088460
https://www.thermofisher.com/order/catalog/product/73405-157846A?SID=srch -srp-73405-157846A
https://www.thermofisher.com/order/catalog/product/43903-157831?SID=srch -srp-43903-157831
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