Protein based therapeutics are rapidly growing, offering a promising approach to fight against cancer, autoimmune diseases, and other conditions. The N-glycosylation of proteins and fusion proteins, play crucial roles in maintaining important therapeutic attributes such as thermal stability, pharmacokinetics (PK), and antibody effector functions. However, the bioreactor environment is variable, and the sialylation period changes throughout upstream process. Therefore, a high-throughput method is necessary to maintain optimal %sialylation while monitoring glycoprotein production. Herein, we utilized Agilent instant-PC-N-glycan® kit to carry out analysis of heavily glycosylated proteins to analyze cell culture fluids (CCFs). Moreover, the results obtained from the CCF samples helped guide immediate decisions to harvest the process during the upstream bioengineering. Later, a RAMAN model was developed by inputting %sialylation of small-scale reactors as a calibration standard and using partial least squared (PLS) + SIMCA (soft indicated modeling by class analogy) combinations to obtain real-time %sialylation of upstream reactors.
Learning Objectives:
Upon completion, participant will be able to understand how the combination of high-throughput N-glycan analysis by instant-PC N-glycan and RAMAN spectroscopy can track the %sialylation in real time.
