Challenges in liquid chromatographic characterization of proteins

•Stability of therapeutic proteins during liquid chromatography is overviewed.•Instrument inertness in terms of protein adsorption and chemical resistance is evaluated.•On-column protein adsorption and recent method development trends are reported.

Various liquid chromatographic techniques are considered standard analytical methods in proteins characterization. These methods provide essential information for drug approval, for biological and life sciences. On the other hand, there are some issues and challenges which have to be taken into account when analyzing these biopharmaceuticals. The aim of this review to summarize the most recent knowledge relating to the following topics: i) sample stability and complexity ii) adsorption problems: instrument inertness iii) adsorption problems: recovery from the stationary phase and iv) challenges in method development. This information is supposed to help practicing chromatographers in the emerging field of therapeutic protein chromatography.