An efficient novel approach is introduced to predict the effectiveness of chemotherapy treatment in lung cancer by monitoring the serum N-glycome of patients combined with artificial intelligence-based data analysis. The study involved thirty-three lung cancer patients undergoing chemotherapy treatments. Serum samples were taken before and after the treatment. The N-linked oligosaccharides were enzymatically released, fluorophore-labeled, and analyzed by capillary electrophoresis with laser-induced fluorescence detection. The resulting electropherograms were thoroughly processed and evaluated by artificial intelligence-based classifiers, i.e., utilizing a machine learning algorithm to categorize the data into two (binary) classes. The classifier analysis method revealed a strong association between the structural changes in the N-glycans and the outcomes of the chemotherapy treatments (ROC >
0.9). This novel combination of bioanalytical and AI methods provided a precise and rapid tool for predicting the effectiveness of chemotherapy.