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Issue 15, 2016
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Complex coacervation of supercharged proteins with polyelectrolytes

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Complexation of proteins with polyelectrolytes or block copolymers can lead to phase separation to generate a coacervate phase or self-assembly of coacervate core micelles. However, many proteins do not coacervate at conditions near neutral pH and physiological ionic strength. Here, protein supercharging is used to systematically explore the effect of protein charge on the complex coacervation with polycations. Four model proteins were anionically supercharged to varying degrees as quantified by mass spectrometry. Proteins phase separated with strong polycations when the ratio of negatively charged residues to positively charged residues on the protein (α) was greater than 1.1–1.2. Efficient partitioning of the protein into the coacervate phase required larger α (1.5–2.0). The preferred charge ratio for coacervation was shifted away from charge symmetry for three of the four model proteins and indicated an excess of positive charge in the coacervate phase. The composition of protein and polymer in the coacervate phase was determined using fluorescently labeled components, revealing that several of the coacervates likely have both induced charging and a macromolecular charge imbalance. The model proteins were also encapsulated in complex coacervate core micelles and micelles formed when the protein charge ratio α was greater than 1.3–1.4. Small angle neutron scattering and transmission electron microscopy showed that the micelles were spherical. The stability of the coacervate phase in both the bulk and micelles improved to increased ionic strength as the net charge on the protein increased. The micelles were also stable to dehydration and elevated temperatures.

Graphical abstract: Complex coacervation of supercharged proteins with polyelectrolytes

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Supplementary files

Article information

01 Jan 2016
11 Feb 2016
First published
11 Mar 2016

Soft Matter, 2016,12, 3570-3581
Article type

Complex coacervation of supercharged proteins with polyelectrolytes

A. C. Obermeyer, C. E. Mills, X. Dong, R. J. Flores and B. D. Olsen, Soft Matter, 2016, 12, 3570
DOI: 10.1039/C6SM00002A

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