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Biomaterials. 2014 Feb;35(7):2295-301. doi: 10.1016/j.biomaterials.2013.11.069. Epub 2013 Dec 17.

Labeling the nucleocapsid of enveloped baculovirus with quantum dots for single-virus tracking.

Author information

1
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China.
2
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, PR China.
3
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, State Key Laboratory of Virology, and Wuhan Institute of Biotechnology, Wuhan University, Wuhan 430072, PR China. Electronic address: dwpang@whu.edu.cn.

Abstract

Utilization of quantum dots (QDs) for single-virus tracking is highly important for understanding virus infection mechanism. However, QD labeling site of real enveloped viruses has been confined to the external envelope so far, causing the impossibility to monitor the late infection events after the loss of envelope. Herein, a strategy to label the internal nucleocapsid of enveloped virus with QDs was proposed. The nucleocapsid of enveloped baculovirus was self-biotinylated during virus replication process in host cells and subsequently labeled with streptavidin-conjugated QDs (SA-QDs). Such host cell-assisted QD labeling was proved to be reliable, specific, efficient and capable of maintaining virus infectivity. Based on such labeling, critical infection events before and after the envelope loss were monitored in real time, including single virus interacting with late endosomes and the subsequent nucleocapsid transporting into cell nucleus. Thus our established QD labeling of enveloped virus nucleocapsid with QDs enables the comprehensive single-virus tracking for deeply understanding virus infection mechanism.

KEYWORDS:

Baculovirus; Biotinylation; Labeling; Nucleocapsid; Quantum dots; Single-virus tracking

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