Using a Microfluidics System to Reproducibly Synthesize Protein Nanoparticles: Factors Contributing to Size, Homogeneity, and Stability

Abstract

The synthesis of Zein nanoparticles (NPs) using conventional methods, such as emulsion solvent diffusion and emulsion solvent evaporation, is often unreliable in replicating particle size and polydispersity between batch-to-batch syntheses. We have systematically examined the parameters for reproducibly synthesizing Zein NPs using a Y-junction microfluidics chip with staggered herringbone micromixers. Our results indicate that the total flow rate of the fluidics system, relative flow rate of the aqueous and organic phase, concentration of the base material and solvent, and properties of the solvent influence the polydispersity and size of the NPs. Trends such as increasing the total flow rate and relative flow rate lead to a decrease in Zein NP size, while increasing the ethanol and Zein concentration lead to an increase in Zein NP size. The solvent property that was found to impact the size of the Zein NPs formed the most was their hydropathy. Solvents that had a hydropathy index most similar to that of Zein formed the smallest Zein NPs. Synthesis consistency was confirmed within and between sample batches. Stabilizing agents, such as sodium caseinate, Tween 80, and Pluronic F-68, were incorporated using the microfluidics system, necessary for in vitro and in vivo use, into Zein-based NPs.

Gold–Protein Composite Nanoparticles for Enhanced X-ray Interactions: A Potential Formulation for Triggered Release

“…Following the procedure to synthesize the ZNPs and ZAuNPs (depicted in Figure 1 a), the final product was analyzed using DLS, UV–vis, and EM ( Figure 1 b and Figure 2 ). ZNPs were optimized formerly for the microfluidics system as described previously [ 8 ]. Briefly, the total flow rate, relative flow rate, solvent concentration and type, and protein concentration were optimized to achieve sub-100 nm nanoparticles reproducibly.…”

“…Zein was purified before use as previously reported by our lab [ 8 ]. Briefly, Zein (0.10 g) was suspended in 15 mL of anhydrous EtOH.…”

“…Zein nanoparticle (ZNP) and Zein–gold hybrid nanoparticle (ZAuNP) synthesis procedures were adapted from a protocol previously reported by our lab [ 8 ]. Briefly, the purified Zein (0.10 g) that was dissolved in 10 mL of 60% ( v / v ) EtOH and filtered, as detailed in Section 2.3 , was subsequently loaded into a syringe (1.0 mL) as the organic phase in the first inlet channel in the microfluidics device.…”

Liposomes: Advancements and innovation in the manufacturing process

“…For example, size-tuned production of liposomes (from 40 to 500 nm depending on the formulation and process parameters e.g. [17,138,148]) with low polydispersity (often below <0.1 PDI) can be achieved through the control of process parameters. The use of microfluidics has also been shown to promotes higher loading within liposomes compared to other commonly used methods; for example, a forty-fold increase in iron oxide loading was achieved using a T mixer compared to other methods [143].…”

Lab-On-A-Chip for the Development of Pro-/Anti-Angiogenic Nanomedicines to Treat Brain Diseases

“…and are used to rapidly synthesize therapeutic NPs in the desired size range, shape and composition [64,98]. Recently, LOCs for producing monodispersed liposomes [99], chitosan NPs [100], protein NPs [101], lipid NPs [102] and other organic NPs [103], some of them with higher EE, were reported [99,102,103].…”