Nanotechnology is one of the most promising technologies of the 21st century, and it is now presenting an enormous impact on target drug delivery. In this context, the recent use of natural vesicle‐like nanoparticles such as extracellular vesicles (i.e., exosomes, microvesicles, and apoptotic bodies) and virus‐like particles is rendering encouraging results mostly because these delivery systems present cargo versatility, favorable body circulating advantages, biocompatibility, immunogenicity, and the capacity to be modified superficially to increase their affinity to a certain target or to control their entrance to the cell. However, some of the biggest challenges toward their clinical implementation are poorly standardized processing operations due to their inherent heterogeneity and expensive, long‐lasting, and difficult to scale isolation procedures that can also affect the stability of the particles. Under these circumstances, chromatographic procedures represent an attractive and favorable alternative to overcome their downstream processing. Moreover, even when standardized chromatographic purification protocols are still in development, great achievements have been made using size exclusion, ionic exchange, hydrophobic interaction, and affinity protocols, mostly because of the correct harnessing of the nanovesicle membrane properties. In this sense, this review focuses on presenting the current understanding on the most promising therapeutic biological nanoparticles and the chromatographic isolation approaches employed in their recovery, providing at the same time recent findings and a general overview of the aspects that might impact the outcome of chromatographic techniques for this application.

The natural triterpene pristimerin possesses interesting potential against several types of cancer. However, its production at the industrial level requires novel environmentally friendly and efficient extraction procedures. Here, we analyze an ethanol–phosphate aqueous two-phase system and the effect of its design parameters such as the tie-line length (TLL) and volume ratio (V R), on the direct extraction of pristimerin. The results showed that a high TLL value of 70% (w/w) and a low V R value of 0.33 allow the obtention of optimal yield and purity values. Partition experiments provided a compound K P value of 3.12 ± 1.21, reaffirming its preference for the ethanol-rich phase. Besides, green assessment positioned the extraction in classification A with 90.93 of 100 points. Furthermore, analysis at different laboratory scales (1×, 10×, and 100×) demonstrated scalability and robustness. Finally, a complete bioprocess based on the obtained results is suggested as a tool to address the pharmaceutical potential of this molecule sustainably and efficiently.