Protein–polyelectrolyte complexes to improve the biological activity of proteins in layer-by-layer assemblies

Straeten, Aurélien vander; Bratek‐Skicki, Anna; Germain, Loïc; D'Haese, Cécile; Eloy, Pierre; Fustin, Charles‐André; Dupont-Gillain, Christine

doi:10.1039/c7nr04345g

Cited by 32 publications

(43 citation statements)

References 34 publications

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“…17 Recently, we brought back to the forefront the idea of premixing enzymes with PEs prior to their LbL assembly. 26 It was shown that lysozyme charge can be tuned by complexation in solution with poly(styrenesulfonate) (PSS) and then used as a building block for LbL film constructions. The obtained coatings show a higher hydration level and a higher specific enzymatic activity compared to bare protein molecules assembled with PSS.…”

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confidence: 99%

Integrating Proteins in Layer-by-Layer Assemblies Independently of their Electrical Charge

et al. 2018

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Layer-by-layer (LbL) assembly is an attractive method for protein immobilization at interfaces, a much wanted step for biotechnologies and biomedicine. Integrating proteins in LbL thin films is however very challenging due to their low conformational entropy, heterogeneous spatial distribution of charges, and polyampholyte nature. Protein-polyelectrolyte complexes (PPCs) are promising building blocks for LbL construction owing to their standardized charge and polyelectrolyte (PE) corona. In this work, lysozyme was complexed with poly(styrenesulfonate) (PSS) at different ionic strengths and pH values. The PPCs size and electrical properties were investigated, and the forces driving complexation were elucidated, in the light of computations of polyelectrolyte conformation, with a view to further unravel LbL construction mechanisms. Quartz crystal microbalance and atomic force microscopy were used to monitor the integration of PPCs compared to the one of bare protein molecules in LbL assemblies, and colorimetric assays were performed to determine the protein amount in the thin films. Layers built with PPCs show higher protein contents and hydration levels. Very importantly, the results also show that LbL construction with PPCs mainly relies on standard PE-PE interactions, independent of the charge state of the protein, in contrast to classical bare protein assembly with PEs. This considerably simplifies the incorporation of proteins in multilayers, which will be beneficial for biosensing, heterogeneous biocatalysis, biotechnologies, and medical applications that require active proteins at interfaces.

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Integrating Proteins in Layer-by-Layer Assemblies Independently of their Electrical Charge

et al. 2018

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“…[ 7,8 ] Recently the multilayer formation between polyelectrolytes and protein/polyelectrolyte complexes was proposed as a method for stable multilayers. [ 9,10 ] PEMLs are proposed for applications related to tissue engineering, [ 11,12 ] drug delivery [ 13 ] and cell culture in general. [ 14–17 ] LbL self‐assembled PEMLs have effectively maintained the concentration of a growth factor for cell cultures to avoid repeated growth factor addition and intervention.…”

Section: Introductionmentioning

confidence: 99%

Polysaccharide–Protein Multilayers Based on Chitosan–Fibrinogen Assemblies for Cardiac Cell Engineering

Κιτσαρά

Tassis

Papagiannopoulos

et al. 2021

Macromolecular Bioscience

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The cell and tissue culture substrates play a pivotal role in the regulation of cell‐matrix and cell‐cell interactions. The surface properties of the materials control a wide variety of cell functions. Amongst various methods, layer‐by‐layer (LbL) assembly is a versatile surface coating technique for creating controllable bio‐coatings. Here, polysaccharide/protein multilayers are proposed, which are fabricated by immersive LbL assembly and based on the chitosan/fibrinogen pair for improving the adhesion and spreading of cardiomyocytes. Two approaches in LbL assembly are employed for clarifying the effect of the bilayers order and their concentration on cardiomyocytes viability and morphology. Fourier transform infrared spectroscopy (FTIR) measurements show that the adsorption of the biopolymers is enhanced during the LbL deposition in a synergistic manner. Contact angle measurements indicate that the multilayers are alternating from less to more hydrophilic behavior depending on the biopolymer that is added last. Confocal microscopy with immunostained fibrinogen reveals that the amount of the protein is higher when the concentration of the immersion solution is increased, however, for low solution concentration it is speculated that interdigitation between the separate biopolymer layers takes place. This work motivates the use of fibrinogen in polysaccharide/protein multilayers for enhanced cytocompatibility in cardiac tissue engineering.

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“…Another technique that has recently gained interest to stabilize proteins is their complexation with polyelectrolytes (62). The formation of protein-polyelectrolyte complexes is a self-assembly process known for more than a decade (63,64) that is now used in drug formulation (65)(66)(67) or in layer-by-layer deposition methods (62). The spontaneous assembly of enzyme-polyelectrolyte complexes (EPCs) is directed by electrostatic and van der Waals forces (68,69).…”

Section: Introductionmentioning

confidence: 99%

“…Generally speaking, enzyme stability can be improved through immobilization on a support (56), through the formation of cross-linked enzyme aggregates (CLEAs) (57) or through the creation of enzyme mutants by directed evolution (58)(59)(60)(61). Another technique that has recently gained interest to stabilize proteins is their complexation with polyelectrolytes (62). The formation of protein-polyelectrolyte complexes is a self-assembly process known for more than a decade (63,64) that is now used in drug formulation (65)(66)(67) or in layer-by-layer deposition methods (62).…”

Section: Introductionmentioning

confidence: 99%

Hybrid Chemo-Biocatalysts Prepared in One Step from Zeolite Nanocrystals and Enzyme-Polyelectrolyte Complexes

Verren¹,

Smeets²,

Straeten³

et al. 2020

Preprint

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The combination of heterogeneous catalysts and enzymes, in so-called hybrid catalysts, is an attractive strategy to effectively run chemoenzymatic reactions. Yet, the preparation of such bifunctional materials remains challenging because both the inorganic and the biological moieties must be integrated in the same solid, while preserving their intrinsic activity. Combining an enzyme and a zeolite, for example, is complicated because the pores of the zeolite are too small to accommodate the enzyme and a covalent anchorage on the surface is often ineffective. Herein, we developed a new pathway to prepare a hybrid catalyst built from glucose oxidase and TS-1 zeolite. Such hybrid material can catalyze the in situ formation of H2O2, which is subsequently used by the zeolite to trigger the epoxidation of allylic alcohol. Starting from an enzymatic solution and a suspension of zeolite nanocrystals, the hybrid catalyst is obtained in one step, using a continuous spray drying method. While enzymes are expectedly unable to resist the conditions used in spray drying (temperature, shear stress, etc.), we leverage on the preparation of “enzyme-polyelectrolyte complexes” (EPCs) to increase the enzyme stability. Importantly, the use of EPCs also appears to prevent enzyme leaching and to stabilize the enzyme against pH changes. We show that the one-pot preparation by spray drying gives access to hybrid catalysts with unprecedented performance in the targeted chemoenzymatic reaction. Interestingly, the hybrid catalyst performs much better than the two catalysts operating as separate entities. We anticipate that this strategy could be used as an adaptable method to prepare other types of multifunctional materials.<br>

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Protein–polyelectrolyte complexes to improve the biological activity of proteins in layer-by-layer assemblies

Cited by 32 publications

References 34 publications

Integrating Proteins in Layer-by-Layer Assemblies Independently of their Electrical Charge

Integrating Proteins in Layer-by-Layer Assemblies Independently of their Electrical Charge

Polysaccharide–Protein Multilayers Based on Chitosan–Fibrinogen Assemblies for Cardiac Cell Engineering

Hybrid Chemo-Biocatalysts Prepared in One Step from Zeolite Nanocrystals and Enzyme-Polyelectrolyte Complexes

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