Polyglycidol-based nanohydrogels (nHGs) have been prepared by optimizing the use of liposome master templates resulting in a high-yielding and more practical one-pot process to provide materials capable of carrying drugs of adverse chemical nature. The nanogels prepared with the one-pot method showed favorable kinetics for the release of either Nile Red (NR) or lysozyme (LYS), loaded with gel precursors such as semi-branched poly(glycidol allylglycidyl ether), PEG dithiol (1KDa), a free radical initiator and liposomal lipids at the liposome formation step. This process is superior to a comparable step-wise traditional approach and circumvents loading of the gel precursors with the hydrophilic drug into preformed liposome templates. A thiol-ene crosslinking reaction accomplishes the formation of the nanonetwork resulting in nHGs prepared in the traditional step-wise (nHG-SW) approach and the one-pot (nHG-OP) process. Both nanogel networks were characterized in terms of particle size and zeta (ζ) potential with average values of 148nm±39nm and -25.9mV±9.2 for the nHG-SW and 132nm±32 and -23.1mV±9.7 for the nHG-OPs. Loading efficiency for both of the nanogels with NR was determined by spectrophotometry to be 28% (nHP-SW) and 31% (nHP-OP). The LYS loading was based on the target loading of 10μg/mg for both nanogels found to be 84% and 86% for the nHG-SW and nHP-OP, respectively. As proof of concept for combination drug delivery, the in vitro release of both drug mimics, NR and LYS, were monitored under physiologically relevant conditions by an optimized dialysis method. The implementation of the multi-functional and semi-branched polyglycidol is recognized as the main contributor for the observed highly controlled release of proteins that are otherwise rapidly released from common PEG-based nanogel networks. Furthermore, the one-pot process led to be the most favorable drug delivery system based on the release kinetics pointing to a denser polymer network.