Trypsin-, proteinase K-, and ribonuclease T1-functionalized magnetic nanoparticle-immobilized enzyme reactors based on polyelectrolyte multilayer platform for biopharmaceutical analysis

) of test substrates and compared to the corresponding enzyme in solution. The amount of bound enzyme was quantified by Micro BCA assay measuring the absorbance at 562 nm. Furthermore, the developed IMERs were applied to protein and RNA digestion, and the resulting fragments were analyzed by HPLC-ESI-QTOF-MS/MS, demonstrating their applicability for biomolecular analysis. Finally, successful mRNA digestion isolated from a vaccine documented the practical applicability of the RNase T1 IMER and the suitability of the polyelectrolyte multilayer enzyme immobilization platform.