2 research outputs found

    Production of nano-protein particles in living cells

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    The escalating demand for therapeutic proteins calls for advanced biomanufacturing and formulation strategiesto ensure efficient production and timely supply. Productions of therapeutic proteins, as well as many otherproteins, require high purity and stability. However, the manufacturing process is bottlenecked by the purifi-cation process. We have established a novel platform leveraging the capabilities of living E. coli cells as cellfactory, for efficient synthesis (protein expression) and easy separation (protein particle formation) of a modelprotein, Red Fluorescent Protein (mRFP). With the designed plasmid, the target protein (mRFP) was heterolo-gously expressed inside cells, and directed to form protein particles, and the red fluorescence of these particlesproved their correct folding and bioavailability. Target protein production was validated through SDS-PAGE andfluorescent microscopy. The cells with protein particles and the isolated protein particles were characterized bySEM, optical and fluorescent microscopic. The new cell factory technology developed for mRFP production hasexhibited broad potential in advancing biomanufacturing, which can be further developed to enable the efficientbioproduction of other therapeutic proteins and peptides.</p

    Positive and negative effects of graphite flake and monolayer graphene oxide templates on protein crystallization

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    Heterogeneous template-induced nucleation is a promising way to regulate protein crystallization events and could be employed for purification processes and crystallographic studies. Protein crystallization process with graphite and graphene oxide, as heterogeneous templates, were investigated. More than 640 hanging drops with different concentrations of Lysozyme (30, 50, 70, 100 mg/mL) and NaCl (0.7, 0.9, 1.1, 1.3, 1.5 M) were crystallised at 4 °C with or without graphite/graphene oxide templates. The induction times and crystallization process were observed under the microscope. The lysozyme in the solutions with graphite flakes nucleated faster under all the conditions than the lysozyme with equal experimental conditions without templates. The crystals preferred to grow around the edge of graphite flakes than on the flat surfaces. In the droplets with monolayer graphene oxide, more crystals appeared around graphene oxide particles, and the faster or slower nucleation processes with templates were dependent on the lysozyme and NaCl concentrations. Graphene oxide templates strongly inhibited nucleation at high lysozyme concentrations but promoted nucleation at low lysozyme concentrations. Both heterogeneous templates changed the crystal morphology and the crystallization kinetics. More crystals were observed in the solution with graphite templatesthan with graphene oxide templates and without any template
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