Monolithic columns allow fast, efficient, and reproducible separation of biomolecules such as proteins, plasmid DNA, antibodies, and viral particles, from samples taken at various stages during a bioprocess. Choosing a monolithic column involves considering the desired column volume and length, the flow rate through the column, construction of the housing, and interface to the HPLC equipment. Monolithic columns are used in the pharmaceutical process development for reaction monitoring, column fraction screening, and analysis of mother liquors and unstable analytes.