Glycosylation is a crucial quality attribute for protein therapeutics, demanding close monitoring. While established assays like HILIC-FLD and LC-MS are effective for glycoproteins with few glycosylation sites, they are less suitable for those with multiple sites, leading to complex, time-consuming data analysis. This study evaluates high-performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) as a faster alternative for monitoring glycosylation in SARS-CoV-2 spike protein production batches. Using glycoengineered spike proteins, HPAEC-PAD accurately and reproducibly detected major and minor glycosylation differences, aligning well with HILIC-FLD and LC-MS results. This suggests HPAEC-PAD offers a rapid, robust method for glycosylation monitoring in complex biotherapeutics.

Janelle Sauvageau, Izel Koyuturk, Frank St. Michael, Denis Brochu, Marie-France Goneau, Ian Schoenhofen, Sylvie Perret, Alexandra Star, Anna Robotham, Arsalan Haqqani, John Kelly, Michel Gilbert, Yves Durocher