Advances in mass spectrometry instrumentation and experimental design have led to significant inroads in the characterization of biological molecules, thus translating to new applications in the field of lipidomics, biotherapeutics, proteomics and structural biology. Ultraviolet photodissociation (UVPD) uses UV laser pulses to activate ions, resulting in extensive fragmentation of many classes of molecules. UVPD offers a versatile MS/MS technology for characterization of lipids, nucleic acids, peptides and proteins,. Owing to the fast, high energy deposition of UV photoactivation, products may retain non-covalently bound ligands or labile modifications, allowing localization of ligand binding sites or modifications. There has also been growing interest in employing MS/MS strategies to examine native protein structures by disassembling the complexes and sequencing the constituent proteins in the gas phase. This presentation will highlight the capabilities of UVPD for characterization of biological molecules and cover new strategies to extend the performance of UVPD, such as integrating UVPD with proton transfer charge reduction, gas-phase fractionation, and ion mobility and implementation of hybrid MS3 methods.