Post-Translational Modification of Bionanoparticles as a Modular Platform for Biosensor Assembly

Sun, Qing; Chen, Qi; Blackstock, Daniel; Chen, Wilfred

doi:10.1021/acsnano.5b03688

Cited by 47 publications

(56 citation statements)

References 45 publications

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“…Another class of PNPs that is gathering more attention is based on multimeric enzymes that naturally self-assemble into nanocages of varying sizes [20–22]. These nanostructures provide naturally-evolved, confined reaction spaces for enhanced cellular metabolism and are ideal candidates as nanoreactors.…”

Section: Enzyme-derived Pnpsmentioning

confidence: 99%

“…E2 is composed of 60 identical monomers that self-assemble into a highly stable cage-like structure with an outer diameter of ~24 nm [23]. The thermostability of E2 nanocages allows for simple purification by heating to 70°C to denature cellular proteins without affecting the E2 assembly (Figure 1c) [22]. …”

Section: Enzyme-derived Pnpsmentioning

confidence: 99%

“…Sortase A from Staphylococcus aureus is often used for its tail-to-head ligations between a small C-terminal LPXTG motif and small N-terminal poly-glycine motif [47]. Sortase A has been utilized for the functionalization of a triglycine modified E2 protein nanocage [6,22]. Our group demonstrated the ligation of LPETG modified elastin-like polypeptide (ELP) and a tetrameric β-galactosidase onto E2 [6].…”

Section: Functionalization Of Protein Nanoparticlesmentioning

confidence: 99%

“…Three LPETG-tagged components, an ELP purification tag, an IgG binding Z-domain, and a nanoluciferase, can be sequentially ligated onto the E2 surface. By controlling the ratios of the reacting LPETG tagged components, the number ligated to E2 can be easily controlled [22]. …”

Section: Functionalization Of Protein Nanoparticlesmentioning

confidence: 99%

“…PNPs are ideally suited for sensing and imaging applications as they provide a high surface-area-to-volume ratio [57] and the potential to be simultaneously modified in both the interior and exterior [58] for multifunctional capabilities [22]. Functional molecules such as fluorophores, metal nanoparticles, antibodies, and aptamers have been displayed on the PNP surface to create new functionalities [22,38,58,59].…”

Section: Biosensing and Bioimaging Applications Of Pnpsmentioning

confidence: 99%

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Protein nanoparticles as multifunctional biocatalysts and health assessment sensors

Raeeszadeh‐Sarmazdeh

Hartzell

Price

et al. 2016

Current Opinion in Chemical Engineering

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The use of protein nanoparticles for biosensing, biocatalysis and drug delivery has exploded in the last few years. The ability of protein nanoparticles to self-assemble into predictable, monodisperse structures is of tremendous value. The unique properties of protein nanoparticles such as high stability, and biocompatibility, along with the potential to modify them led to development of novel bioengineering tools. Together, the ability to control the interior loading and external functionalities of protein nanoparticles makes them intriguing nanodevices. This review will focus on a number of recent examples of protein nanoparticles that have been engineered towards imparting the particles with biocatalytic or biosensing functionality.

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Section: Enzyme-derived Pnpsmentioning

confidence: 99%

Section: Enzyme-derived Pnpsmentioning

confidence: 99%

Section: Functionalization Of Protein Nanoparticlesmentioning

confidence: 99%

Section: Functionalization Of Protein Nanoparticlesmentioning

confidence: 99%

Section: Biosensing and Bioimaging Applications Of Pnpsmentioning

confidence: 99%

See 3 more Smart Citations

Protein nanoparticles as multifunctional biocatalysts and health assessment sensors

Raeeszadeh‐Sarmazdeh

Hartzell

Price

et al. 2016

Current Opinion in Chemical Engineering

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Polymeric SpyCatcher Scaffold Enables Bioconjugation in a Ratio‐Controllable Manner

Jia

Minamihata

Ichinose

et al. 2017

Biotechnology Journal

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Conjugating enzymes into a large protein assembly often results in an enhancement of overall catalytic activity, especially when different types of enzymes that work cooperatively are assembled together. However, exploring the proper method to achieve protein assemblies with high stability and also to avoid loss of the function of each component for efficient enzyme clustering is remained challenging. Assembling proteins onto synthetic scaffolds through varied post-translational modification methods is particularly favored since the proteins can be site-specifically conjugated together with less activity loss. Here, a SpyCatcher polymer is prepared through catalytic reaction of horseradish peroxidase (HRP) and serves as a polymeric proteinaceous scaffold for construction of protein assemblies. Taking advantage of the favorable SpyCatcher-SpyTag interaction, SpyTagged proteins can be easily assembled onto the polymeric SpyCatcher scaffold with controllable binding ratio and site specificity. Firstly, the feasibility of construction of ratio-controllable binary artificial hemicellulosomes by assembling endoxylanase and arabinofuranosidase is explored. This construct achieves higher sugar conversion than that of the free enzymes when the proportion of arabinofuranosidase is high, because the close spatial proximity of the enzymes allows them to work in a synergistic manner. Another application for biosensing is developed by conjugating SpyTagged Nanoluc and protein G onto SpyCatcher polymer. Due to the protein clustering effect, an amplified luminescent intensity is achieved by the resulting conjugates than chimera protein of Nanoluc and protein G in ovalbumin detection in ELISA.

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One‐step affinity capture and precipitation for improved purification of an industrial monoclonal antibody using Z‐ELP functionalized nanocages

Swartz¹,

Traylor

et al. 2017

Biotech & Bioengineering

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Protein A chromatography has been identified as a potential bottleneck in the monoclonal antibody production platform, leading to increased interest in non-chromatographic capture technologies. Affinity precipitation using environmentally responsive, Z-domain-elastin-like polypeptide (Z-ELP) fusion proteins has been shown to be a promising alternative. However, elevated temperature and salt concentrations necessary for precipitation resulted in decreased antibody monomer content and reduced purification capacity. To improve upon the existing technology, we reported an enhanced affinity precipitation of antibodies by conjugating Z-ELP to a 25 nm diameter, self-assembled E2 protein nanocage (Z-ELP-E2). The enlarged scale of aggregate formation and IgG-triggered crosslinking through multi-valent binding significantly outperformed traditional Z-ELP-based methods. In the current work, we sought to develop an affinity precipitation process capable of purifying industrial monoclonal antibodies (mAbs) at ambient temperature with minimal added salt. We discovered that the mAb-nanocage complex aggregated within 10 min at room temperature without the addition of salt due to the enhanced multi-valent cross-linking. After precipitating out of solution, the complex remained insoluble under all wash buffers tested, and only resolubilized after a low pH elution. Through optimization of key process steps, the affinity precipitation yield and impurity clearance met or exceeded protein A chromatography performance with 95% yield, 3.7 logs host cell protein reduction, and >5 logs of DNA reduction from mAb cell culture. Because of the operational flexibility afforded by this one-step affinity capture and precipitation process, the Z-ELP-E2 based approach has the potential to be a viable alternative to platform mAb purification.

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Post-Translational Modification of Bionanoparticles as a Modular Platform for Biosensor Assembly

Cited by 47 publications

References 45 publications

Protein nanoparticles as multifunctional biocatalysts and health assessment sensors

Protein nanoparticles as multifunctional biocatalysts and health assessment sensors

Polymeric SpyCatcher Scaffold Enables Bioconjugation in a Ratio‐Controllable Manner

One‐step affinity capture and precipitation for improved purification of an industrial monoclonal antibody using Z‐ELP functionalized nanocages

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