Fluorescence excitation-emission matrix (EEM) spectroscopy is used for quantitative predictive analysis of glycoprotein production in a CHO cell fed-batch process. EEM spectra of complex solutions are very sensitive to compositional change and as cultivation progressed, the emission of tyrosine, tryptophan, and the glycoprotein product showed significant differences, and this was used to follow culture progress via chemometrics. This clearly showed the increase of the unique dityrosine emission from the product glycoprotein as the process progressed. A second aspect of the study involved developing quantitative predictive models of process performance based on glycoprotein yield. Accurate predictions with relative errors of between <5% were obtained for samples extracted from the 100 to 5000L scale bioreactors. This methodology opens the possibility of early stage intervention for poorly performing lots.