Modern chromatographic and mass spectrometric techniques for protein biopharmaceutical characterization

• Protein biopharmaceuticals are highly complex and heterogeneous macromolecules.
• Chromatography and MS are at the forefront in protein characterization.
• Primary and higher order structures and pharmacokinetic properties can be assessed.

Protein biopharmaceuticals such as monoclonal antibodies and therapeutic proteins are currently in widespread use for the treatment of various life-threatening diseases including cancer, autoimmune disorders, diabetes and anemia. The complexity of protein therapeutics is far exceeding that of small molecule drugs; hence, unraveling this complexity represents an analytical challenge. The current review provides the reader with state-of-the-art chromatographic and mass spectrometric tools available to dissect primary and higher order structures, post-translational modifications, purity and impurity profiles and pharmacokinetic properties of protein therapeutics.