Issue link: https://resources.nanotempertech.com/i/1050512
Finally, the N-linked glycan profiles were acquired for the samples and compared to the reference. Again, an appropriate algorithm of UGA Biopharma was used to rank the samples (Table 3). Table 3. Samples ranked according to the N-linked glycan similarity index. Rank SampleID 1 15 2 11 3 09 4 06b 5 03 6 07 7 05 To compare the results obtained by a single method (nanoDSF thermal profiling) and more specialized analyses used in biosimilar development, we combined the ranks of charge variant profiling, sialic acid quantification and N-linked glycan profiles into a combined similarity rank (Table 4). Table 4. Samples ranked according to the combined charge variants, sialic acid content and N-linked glycan similarity index. Combined rank SampleID 1 11 2 09 3 03 3 15 4 06b 5 07 6 05 Both approaches for biosimilar candidate selection, nanoDSF profiling and three combined HPLC methods identified sample #11 as the one showing the best match to the reference product (Figure 4). In addition, there was an overlap for 5 out of 7 samples with most similarity for the two ranking strategies. The comparison shows an excellent agreement between the similarity ranking obtained by combined charge variants, N-linked glycans and sialic acid content methods with the nanoDSF thermal unfolding profile ranking. Figure 4: N-linked glycan analysis of the reference product and candidate 014-11. Conclusions This study shows that nanoDSF has the potential to revolutionize similarity screening processes in biosimilar profiling. The best-in- class precision, reproducibility and data quality of the Prometheus series delivers high-quality thermal unfolding profiles which show even smallest differences between biosimilar variants. Up to 48 samples, each with a volume of just 10 µl and less than 1 µg of protein, can be measured in parallel. nanoDSF is therefore orders of magnitude faster than the complimentary screening methods used here, which require ~ 1 week of lab work. Moreover, 150-times less protein is used for the analysis, rendering nanoDSF the perfect tool for large- scale screenings at early stages of biosimilar development. The optional automation allows for unattended analysis of hundreds of samples per day, resulting in a yet unknown productivity. The flexibility, low consumable costs and maintenance-free operation of the system allows for its integration into virtually every step in the biosimilar development process. This enables scientists to perform quick and meaningful screenings with minimal time and material requirements to filter candidates with similar structural properties based on their unfolding patterns.