The negative effect that a RAFT agent with a poor leaving group has on the evolution of molecular dispersity in a RAFT-mediated polymerization was shown to be mitigated by performing the polymerization in semi-batch mode.
Modified chitosan and modified poly(styrene-alt-maleic anhydride) (SMA) were synthesized and utilized to coat superparamagnetic magnetite nanoparticles (SPMNs) to act as potential Mycobacterium tuberculosis (Mtb) capturing substrates. Chitosan and SMA were modified to form quaternary derivatives which have the potential to interact with the Mtb cell wall. The nano-substrates were also surface functionalized with a carbohydrate-binding protein, namely Concanavalin A (Con A), which can bind to the Mtb cell wall. Chitosan coated SPMNs were synthesized by in situ co-precipitating Fe 2+ and Fe 3+ with chitosan followed by further modification. SMA coated SPMNs were synthesized by activating the iron oxide core with 3-aminopropyl(triethoxysilane) followed by further modification. The nano-substrates were evaluated for their affinity and thus capturing capabilities utilizing the mCherry fluorophore tagged bacillus Calmette-Guérin (BCG) strain of Mycobacterium bovis, a live attenuated Mtb-mimic. Quaternary SMA (SMI-qC 12 ) revealed the highest overall affinity for BCG-mCherry (through electrostatic and hydrophobic interactions) whereas Con A immobilized chitosan had the highest affinity for the chitosan coated nanoparticles. The SPMNs were coated with three different polymer loadings and a dilution study was performed to determine the limit of detection. The 0.9 g loaded SMI-qC 12 SPMNs revealed the highest affinity for BCG-mCherry as determined via fluorescence microscopy and transmission electron microscopy.
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