Protein separation by sequential selective complex coacervation

Zhou, Jin; Cai, Ying; Wan, Yat-wah; Wu, Bohang; Wang, Zhibin; Zhang, Xinxin; Hu, Weiwei; Stuart, Martien A. Cohen; Wang, Junyou

doi:10.1016/j.jcis.2023.06.119

Cited by 10 publications

(6 citation statements)

References 53 publications

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“…30,31 The branched structure of PEI causes steric hindrance, which makes the internal amines less accessible than the external amines, thus a slight excess of PEI is required to achieve charge stoichiometry. 27 Further increasing PEI leads to a decay in intensity, which may be from the dissociation of the formed assemblies, as found in previous studies. 32,33 Correspondingly, the particle size also exhibits an increasing and then decreasing change with increasing f +/À , and at the PMC the particles display a hydrodynamic radius (R h ) of B34.6 nm (Fig.…”

Section: Preparation Of Pic Vesiclessupporting

confidence: 73%

“…In this study, we selected branched poly(ethylenimine) (PEI) as the cationic polymer (Scheme 1a), which bears primary, secondary and tertiary amines with a ratio of 1 : 2 : 1, and p K a values of 10.5, 7.5–8.5, and 5.0, respectively. 27 The polyion-neutral diblock copolymer is PSS 96 - b -PEO 113 , which contains sulfonate groups, which are strong ionic groups with negative charges independently of pH. The charge-driven assembly occurs in MES buffer at pH 6.0, thus the tertiary amines in PEI are hardly protonated, while the primary and secondary amines are protonated completely, giving 75% of amines with positive charges.…”

Section: Resultsmentioning

confidence: 99%

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Stable and permeable polyion complex vesicles designed as enzymatic nanoreactors

Wan,

Qiu,

Zhou

et al. 2024

Soft Matter

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Section: Preparation Of Pic Vesiclessupporting

confidence: 73%

Section: Resultsmentioning

confidence: 99%

Stable and permeable polyion complex vesicles designed as enzymatic nanoreactors

Wan,

Qiu,

Zhou

et al. 2024

Soft Matter

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“…Increasing NaCl concentration will weaken the electrostatic interaction between the polymer and protein due to the screening effects, thereby inhibiting the formation of the coacervates accompanied by a decay of the maximum turbidity . This screening effects also interrupt the polymer partition, namely, more proteins and polymers may stay in the top dilution phase rather than involved in the bottom coacervates . Nevertheless, the moderate decline of the interaction is favorable for creating more liquid-like coacervate droplets (Figure S5).…”

Section: Resultsmentioning

confidence: 99%

“…49 This screening effects also interrupt the polymer partition, namely, more proteins and polymers may stay in the top dilution phase rather than involved in the bottom coacervates. 50 Nevertheless, the moderate decline of the interaction is favorable for creating more liquid-like coacervate droplets (Figure S5). Of course, further increasing NaCl concentration, e.g., up to 100 mM, the BSA−PDMAEMA 223 interaction is suppressed so much that most of the polymers may be free or form small clusters suspending in solution, which is validated by the low turbidity and microscopic images (Figure 3e,f).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Effects of Control Factors on Protein–Polyelectrolyte Complex Coacervation

Zhou,

Wan,

Cohen Stuart

et al. 2023

Biomacromolecules

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Protein−polyelectrolyte complex coacervation is of particular interest for mimicking intracellular phase separation and organization. Yet, the challenge arises from regulating the coacervation due to the globular structure and anisotropic distributed charges of protein. Herein, we fully investigate the different control factors and reveal their effects on proteinpolyelectrolyte coacervation. We prepared mixtures of BSA (bovine serum albumin) with different cationic polymers, which include linear and branched polyelectrolytes covering different spacer and charge groups, chain lengths, and polymer structures. With BSA-PDMAEMA [poly(N,N-dimethylaminomethyl methacrylate)] as the main investigated pair, we find that the moderate pH and ionic strength are essential for the adequate electrostatic interaction and formation of coacervate droplets. For most BSA−polymer mixtures, excess polyelectrolytes are required to achieve the full complexation, as evidenced by the deviated optimal charge mixing ratios from the charge stoichiometry. Polymers with longer chains or primary amine groups and a branched structure endow a strong electrostatic interaction with BSA and cause a bigger charge ratio deviation associated with the formation of solid-like coacervate complexes. Nevertheless, both the liquid-and solid-like coacervates hardly interrupt the BSA structure and activity, indicating the safe encapsulation of proteins by the coacervation with polyelectrolytes. Our study validates the crucial control of the diverse factors in regulating protein−polyelectrolyte coacervation, and the revealed principles shall be instructive for establishing other protein-based coacervations and boosting their potential applications.

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“…Future efforts will look to understand whether the interactions and effects that drive trends in encapsulation play similar roles in modulating the thermal stability of our virus formulations. This potential to extract, concentrate, purify, ,− and stabilize viruses and other biomacromolecules − could allow for the use of coacervation to enable a new generation of more stable and higher performing biologics, sensors, or biocatalysts.…”

Section: Discussionmentioning

confidence: 99%

Design Rules for the Sequestration of Viruses into Polypeptide Complex Coacervates

Joshi,

Decker,

Zeng

et al. 2023

Biomacromolecules

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Encapsulation is a strategy that has been used to facilitate the delivery and increase the stability of proteins and viruses. Here, we investigate the encapsulation of viruses via complex coacervation, which is a liquid–liquid phase separation resulting from the complexation of oppositely charged polymers. In particular, we utilized polypeptide-based coacervates and explored the effects of peptide chemistry, chain length, charge patterning, and hydrophobicity to better understand the effects of the coacervating polypeptides on virus incorporation. Our study utilized two nonenveloped viruses, porcine parvovirus (PPV) and human rhinovirus (HRV). PPV has a higher charge density than HRV, and they both appear to be relatively hydrophobic. These viruses were compared to characterize how the charge, hydrophobicity, and patterning of chemistry on the surface of the virus capsid affects encapsulation. Consistent with the electrostatic nature of complex coacervation, our results suggest that electrostatic effects associated with the net charge of both the virus and polypeptide dominated the potential for incorporating the virus into a coacervate, with clustering of charges also playing a significant role. Additionally, the hydrophobicity of a virus appears to determine the degree to which increasing the hydrophobicity of the coacervating peptides can enhance virus uptake. Nonintuitive trends in uptake were observed with regard to both charge patterning and polypeptide chain length, with these parameters having a significant effect on the range of coacervate compositions over which virus incorporation was observed. These results provide insights into biophysical mechanisms, where sequence effects can control the uptake of proteins or viruses into biological condensates and provide insights for use in formulation strategies.

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Protein separation by sequential selective complex coacervation

Cited by 10 publications

References 53 publications

Stable and permeable polyion complex vesicles designed as enzymatic nanoreactors

Stable and permeable polyion complex vesicles designed as enzymatic nanoreactors

Effects of Control Factors on Protein–Polyelectrolyte Complex Coacervation

Design Rules for the Sequestration of Viruses into Polypeptide Complex Coacervates

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