Large scale modification of biomolecules using immobilized sortase A from Staphylococcus aureus.

摘要: Recently, sortase A (SrtA) from Staphyloccus aureus moved into the focus of bioscience because its ability to incorporate site specific modifications proteins. The enzyme was mostly used modify target proteins in an analytical scale, study biomolecules their cellular context. In this study, we show applicability SrtA mediated ligation for modification a large scale. Therefore, reaction first optimized using peptides and subsequently new conditions were applied scale biotinylation interleukin-8. Furthermore, established C-terminal immobilization on PEG based resin could demonstrate maintaining enzymatic activity. Immobilized significantly facilitates previous protocols as can be easily recycled. Also, removal excess solution whole washing process is accelerated, centrifugation or filtration techniques instead time-consuming chromatography steps.