Enhanced recombinant protein capture, purity and yield from crude bacterial cell extracts by N-Lauroylsarcosine-assisted affinity chromatography

JOSÉ VICENTE CARRATALÁ; JAN ATIENZA GARRIGA; HECTOR LOPEZ LAGUNA; ESTHER VÁZQUEZ; ANTONIO VILLAVERDE; JULIETA M. SANCHEZ; NEUS FERRER MIRALLES

MICROBIAL CELL FACTORIES

BIOMED CENTRAL LTD

Lugar: Londres; Año: 2023

1475-2859

Background Recombinant proteins cover a wide range of biomedical, biotechnological, and industrial needs.Although there are diverse available protocols for their purification from cell extracts or from culture media, many pro‑teins of interest such as those containing cationic domains are difficult to purify, a fat that results in low yields of thefinal functional product. Unfortunately, this issue prevents the further development and industrial or clinical applica‑tion of these otherwise interesting products.Results Aiming at improving the purification of such difficult proteins, a novel procedure has been developed basedon supplementing crude cell extracts with non-denaturing concentrations of the anionic detergent N-Lauroylsar‑cosine. The incorporation of this simple step in the downstream pipeline results in a substantial improvement of theprotein capture by affinity chromatography, an increase of protein purity and an enhancement of the overall processyield, being the detergent not detectable in the final product.Conclusion By taking this approach, which represents a smart repurposing of N-Lauroylsarcosine applied to proteindownstream, the biological activity of the protein is not affected. Being technologically simple, the N-Lauroylsarco‑sine-assisted protein purification might represent a critical improvement in recombinant protein production withwide applicability, thus smothering the incorporation of promising proteins into the protein market.