Recombinant protein (EGFP-Protein G)-coated PbS quantum dots for<i>in vitro</i>and<i>in vivo</i>dual fluorescence (visible and second-NIR) imaging of breast tumors

Sasaki, Akira; Tsukasaki, Yoshikazu; Komatsuzaki, Akihito; Sakata, Takao; Yasuda, Hidehiro

doi:10.1039/c4nr06480a

Cited by 73 publications

(66 citation statements)

References 49 publications

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“…reported a fluorescent unimolecular micelle probe for in vitro and in vivo cancer imaging; the micelle probe included different types of organic fluorescent dyes having dual emission in the visible and NIR regions ( λ =500–750 nm) . In previous work, we developed fluorescent recombinant protein capped PbS QDs that showed dual emission at λ =515 and 1100 nm . We used a recombinant protein (GST‐EGFP‐GB1; GST=glutathione s‐transferase, EGFP=enhanced green fluorescent protein, GB1=immunoglobulin binding domain of protein G) as a protecting reagent to prepare PbS QDs with dual‐color emission resulting from EGFP and QDs.…”

Section: Introductionsupporting

confidence: 65%

“…For the EGFP‐GB1 protein, first the GST‐EGFP‐GB1 protein was expressed and lysed following the protocol previously reported by Sasaki et al . The lysate was clarified by centrifugation at 20 000 g for 30 min to eliminate cell debris.…”

Section: Methodsmentioning

confidence: 99%

“…To overcome this disadvantage, adaptor proteins such as streptavidin and protein A/G have been used for the functionalization of QDs with antibodies . Recently, we reported that a small adaptor protein, immunoglobulin binding domain (B1), could be used to mediate antibody conjugation for the preparation of antibody‐functionalized QDs .…”

Section: Introductionmentioning

confidence: 98%

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Fluorescent, Recombinant‐Protein‐Conjugated, Near‐Infrared‐Emitting Quantum Dots for in Vitro and in Vivo Dual‐Color Molecular Imaging

Tsuboi

2018

ChemBioChem

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Near‐infrared (NIR)‐emitting fluorescent probes are widely used for molecular imaging at the whole‐body level. However, NIR‐emitting fluorescent probes emitting over λ=700 nm are not suitable for molecular imaging at the cellular level, because most of the conventional fluorescence microscopes have very low optical sensitivity in the NIR region. Thus, to achieve fluorescence imaging at the cellular and whole‐body levels by using single probes, visible and NIR‐emitting dual‐color fluorescent probes are desirable. For dual‐color fluorescence molecular imaging, we synthesized fluorescent, recombinant‐protein‐conjugated, NIR‐emitting quantum dots (QDs), in which the recombinant protein consists of enhanced green fluorescent protein (EGFP) and the immunoglobulin binding domain (B1) of protein G. This dual‐color fluorescent QD probe binds the Fc region of immunoglobulin G (IgG) through its B1 domain at the QD surface and acts as a molecular‐imaging probe at both the cellular and whole‐body levels. In this paper, we present the synthesis of fluorescent, recombinant protein (HisEGFP‐GB1)‐conjugated, NIR‐emitting QDs and their application to the dual‐color molecular imaging of breast cancer cells in vitro and in vivo.

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Section: Introductionsupporting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

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Fluorescent, Recombinant‐Protein‐Conjugated, Near‐Infrared‐Emitting Quantum Dots for in Vitro and in Vivo Dual‐Color Molecular Imaging

Tsuboi

2018

ChemBioChem

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“…Monomeric streptavidin has been used to prevent cross-linking of biotinylated cell-surface proteins but generation of the monomeric streptavidin remains challenging [279]. For antibody conjugation, molecular adaptor proteins such as Protein A/G or Fc Receptor provide unique advantages due to oriented attachment to antibody regions that are not needed for target binding, leaving the target-binding domain extended away from the QD surface, without the need for covalent modification of the antibody [226, 280, 281]. Protein A and G are also oligomers that can bind to multiple proteins, whereas Fc Receptor I is monomeric and binds to a single antibody [226].…”

Section: Attachment To Molecular Targetsmentioning

confidence: 99%

Quantum dot surface engineering: Toward inert fluorophores with compact size and bright, stable emission

Lim

Schleife

et al. 2016

Coordination Chemistry Reviews

109

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The surfaces of colloidal nanocrystals are complex interfaces between solid crystals, coordinating ligands, and liquid solutions. For fluorescent quantum dots, the properties of the surface vastly influence the efficiency of light emission, stability, and physical interactions, and thus determine their sensitivity and specificity when they are used to detect and image biological molecules. But after more than 30 years of study, the surfaces of quantum dots remain poorly understood and continue to be an important subject of both experimental and theoretical research. In this article, we review the physics and chemistry of quantum dot surfaces and describe approaches to engineer optimal fluorescent probes for applications in biomolecular imaging and sensing. We describe the structure and electronic properties of crystalline facets, the chemistry of ligand coordination, and the impact of ligands on optical properties. We further describe recent advances in compact coatings that have significantly improved their properties by providing small hydrodynamic size, high stability and fluorescence efficiency, and minimal nonspecific interactions with cells and biological molecules. While major progress has been made in both basic and applied research, many questions remain in the chemistry and physics of quantum dot surfaces that have hindered key breakthroughs to fully optimize their properties.

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“…[67] Various biological coatings of PbS QDs have been developed, such as glutathione [68] or proteins. [69,70] Hong et al first synthesized Ag 2 SQ Ds in an organic phase anda pplied themt oi nv ivo NIR-I fluorescence imaging ( Figure 11). [71] Chen and Wang reported NIR-II imaging to track stem cells in living animals, with both high spatial and temporal resolution based on Ag 2 SQDs.…”

Section: Quantum Dots (Qds)mentioning

confidence: 99%

Recent Progress in Fluorescence Imaging of the Near‐Infrared II Window

Miao

Zhu

et al. 2018

ChemBioChem

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Near‐infrared (NIR) fluorescent materials are considered to be the most promising labeling reagents for sensitive determination and biological imaging due to the advantages of lower background noise, deeper penetrating capacity, and less destructive effects on the biomatrix over those of UV and visible fluorophores. In the past decade, advances in biomedical fluorescence imaging in the NIR region have focused on the traditional NIR window (NIR‐I; λ=700–900 nm), and have recently been extended to the second NIR window (NIR‐II; λ=1000–1700 nm). In vivo NIR‐II fluorescence imaging outperforms imaging in the NIR‐I window as a result of further reduced absorption, tissue autofluorescence, and scattering. In this review, the applications of four types of NIR‐II fluorescent materials, organic fluorophores, quantum dots, rare‐earth compounds, and single‐walled carbon nanotubes, are summarized and future trends are discussed. Some methods to enhance the NIR‐II fluorescence quantum yield are also proposed.

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Recombinant protein (EGFP-Protein G)-coated PbS quantum dots forin vitroandin vivodual fluorescence (visible and second-NIR) imaging of breast tumors

Abstract: We report a one-step synthetic strategy for the preparation of recombinant protein (EGFP-Protein G)-coated PbS quantum dots for dual (visible and second-NIR) fluorescence imaging of breast tumors at the cellular and whole-body level.

Cited by 73 publications

References 49 publications

Fluorescent, Recombinant‐Protein‐Conjugated, Near‐Infrared‐Emitting Quantum Dots for in Vitro and in Vivo Dual‐Color Molecular Imaging

Fluorescent, Recombinant‐Protein‐Conjugated, Near‐Infrared‐Emitting Quantum Dots for in Vitro and in Vivo Dual‐Color Molecular Imaging

Quantum dot surface engineering: Toward inert fluorophores with compact size and bright, stable emission

Recent Progress in Fluorescence Imaging of the Near‐Infrared II Window

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