Detecting Species-Specific Cytosolic Transgene Protein Supporting Liver Gene Therapy Studies

Cross-species translation of preclinical models is critical for the development of recombinant adeno-associated virus (rAAV) gene therapy. One factor that can make translation difficult is the inability to determine human transgene protein expression alongside endogenous protein levels in preclinical animals with high sequence homology. Disease specific mouse knockout models are commonly utilized in preclinical drug development. Some models such as Pahenu2 mice for Phenylketonuria may only exhibit a loss of enzymatic activity due to a point mutation while retaining normal protein levels. Due to the background expression of cross-reactive protein in preclinical models, it is important to develop a species-specific transgene protein assay to determine the amount of transgene protein being expressed after rAAV administration. This presentation will address the bioanalytical challenges in developing an assay to detect species-specific cytosolic transgene protein to aid in the understanding of the translation of gene therapy in multiple species