In previous studies with cystic fibrosis (CF) IB3-1 lung epithelial cells in culture, we identified 194 unique high abundance proteins by conventional 2DGE and mass spectrometry (Pollard et al, 2005; Reference 13). In the present work we have compared the IB3-1 cells with IB3-1/S9 daughter cells repaired by gene transfer with AAV-[wildtype]CFTR. We report that gene transfer results in significant changes in silver-stain intensity of only 20 of the 194 proteins. However, simultaneous measurement of de novo biosynthetic rates with 35[S]methionine of all 194 proteins in both cell types results in the identification of an additional 31 CF specific proteins. Of the 51 proteins identified by this hybrid approach, only 6 proteins change similarly in both the mass and kinetics categories. This kinetic portion of the high abundance CF proteome, hidden from direct analysis of abundance, includes proteins from transcription and signaling pathways, such as NFB; chaperones such as HSC70; cytoskeletal proteins; and others. Connectivity analysis indicates that ca. 30% of the 51 member hybrid high abundance CF proteome interacts with the NFB signaling pathway. In conclusion, measurement of biosynthetic rates on a global scale can be used to identify disease-specific differences within the high abundance cystic fibrosis proteome. Most of these kinetically defined proteins are unaffected in expression level when using conventional silver-stain analysis. We anticipate that this novel hybrid approach to discovery of the high abundance CF proteome will find general application to other proteomic problems in biology and medicine.