Surface labeling of enveloped virus with polymeric imidazole ligand-capped quantum dots via the metabolic incorporation of phospholipids into host cells

Zhao, Xia; Shen, Yulong; Adogla, Enoch A.; Viswanath, Anand; Benicewicz, Brian C.; Greytak, Andrew B.; Lin, Yuan; Wang, Qian

doi:10.1039/c6tb00263c

Cited by 19 publications

(18 citation statements)

References 43 publications

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“…Unbound viruses and QDs were removed just by washing the cells. Such a strategy can thoroughly evade ultracentrifugation, dialysis, and ultrafiltration processes that are indispensable for removing the cell-derived reactive molecules, redundant functional reagents, unlabeled viruses, or unbound QDs in many other labeling strategies (29)(30)(31)(32)(33)(34). This strategy further minimized and simplified the handling of viruses, making the QD labeling milder and more convenient.…”

Section: Resultsmentioning

confidence: 99%

Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking

Zhang

Wang

et al. 2020

mBio

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Quantum dots (QDs) possess optical properties of superbright fluorescence, excellent photostability, narrow emission spectra, and optional colors. Labeled with QDs, single molecules/viruses can be rapidly and continuously imaged for a long time, providing more detailed information than when labeled with other fluorophores. While they are widely used to label proteins in single-molecule-tracking studies, QDs have rarely been used to study virus infection, mainly due to a lack of accepted labeling strategies. Here, we report a general method to mildly and readily label enveloped viruses with QDs. Lipid-biotin conjugates were used to recognize and mark viral lipid membranes, and streptavidin-QD conjugates were used to light them up. Such a method allowed enveloped viruses to be labeled in 2 h with specificity and efficiency up to 99% and 98%, respectively. The intact morphology and the native infectivity of viruses were preserved. With the aid of this QD labeling method, we lit wild-type and mutant Japanese encephalitis viruses up, tracked their infection in living Vero cells, and found that H144A and Q258A substitutions in the envelope protein did not affect the virus intracellular trafficking. The lipid-specific QD labeling method described in this study provides a handy and practical tool to readily “see” the viruses and follow their infection, facilitating the widespread use of single-virus tracking and the uncovering of complex infection mechanisms. IMPORTANCE Virus infection in host cells is a complex process comprising a large number of dynamic molecular events. Single-virus tracking is a versatile technique to study these events. To perform this technique, viruses must be fluorescently labeled to be visible to fluorescence microscopes. The quantum dot is a kind of fluorescent tag that has many unique optical properties. It has been widely used to label proteins in single-molecule-tracking studies but rarely used to study virus infection, mainly due to the lack of an accepted labeling method. In this study, we developed a lipid-specific method to readily, mildly, specifically, and efficiently label enveloped viruses with quantum dots by recognizing viral envelope lipids with lipid-biotin conjugates and recognizing these lipid-biotin conjugates with streptavidin-quantum dot conjugates. It is not only applicable to normal viruses, but also competent to label the key protein-mutated viruses and the inactivated highly virulent viruses, providing a powerful tool for single-virus tracking.

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Section: Resultsmentioning

confidence: 99%

Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking

Zhang

Wang

et al. 2020

mBio

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“…Hence, incorporating azide moieties to host cell membrane would enable viral particles to be functionalized with azide without direct chemical modification . Herein, we first synthesized azidoethyl‐choline (AE‐Cho), a metabolic precursor that can be effectively incorporated to phospholipid of cell membrane to form azide‐Choline containing phospholipids through intracellular biosynthetic machinery . The chemical structures of AE‐Cho were confirmed using electrospray ionization mass spectrometry (ESI–MS), as shown in Figure S1 in the Supporting Information.…”

Section: Resultsmentioning

confidence: 99%

In Situ Bioorthogonal Metabolic Labeling for Fluorescence Imaging of Virus Infection In Vivo

Pan

et al. 2017

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Optical fluorescence imaging is an important strategy to explore the mechanism of virus-host interaction. However, current fluorescent tag labeling strategies often dampen viral infectivity. The present study explores an in situ fluorescent labeling strategy in order to preserve viral infectivity and precisely monitor viral infection in vivo. In contrast to pre-labeling strategy, mice are first intranasally infected with azide-modified H5N1 pseudotype virus (N -H5N1p), followed by injection of dibenzocyclooctyl (DBCO)-functionalized fluorescence 6 h later. The results show that DBCO dye directly conjugated to N -H5N1p in lung tissues through in vivo bioorthogonal chemistry with high specificity and efficacy. More remarkably, in situ labeling rather than conventional prelabeling strategy effectively preserves viral infectivity and immunogenicity both in vitro and in vivo. Hence, in situ bioorthogonal viral labeling is a promising and reliable strategy for imaging and tracking viral infection in vivo.

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“… 44 This ligand system is representative of a growing family of hydrophilic, multiply-binding polymers that has emerged as an effective route to biocompatible QDs with high colloidal stability. 26 , 46 – 49 The ligand selected contains approximately equal numbers of poly(ethylene glycol) and imidazole side-chains and has a total molecular weight ∼27 kD. When this ligand exchange is conducted on the benchtop in chloroform solvent, we have shown it to provide water-soluble QDs with long term stability and high brightness.…”

Section: Resultsmentioning

confidence: 99%

Gel permeation chromatography as a multifunctional processor for nanocrystal purification and on-column ligand exchange chemistry

et al. 2016

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Surface labeling of enveloped virus with polymeric imidazole ligand-capped quantum dots via the metabolic incorporation of phospholipids into host cells

Abstract: The successful modification of enveloped virus with polymeric imidazole ligand coated quantum dots was achieved by metabolic labeling and the click reaction.

Cited by 19 publications

References 43 publications

Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking

Lipid-Specific Labeling of Enveloped Viruses with Quantum Dots for Single-Virus Tracking

In Situ Bioorthogonal Metabolic Labeling for Fluorescence Imaging of Virus Infection In Vivo

Gel permeation chromatography as a multifunctional processor for nanocrystal purification and on-column ligand exchange chemistry

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