Impact of H‐aggregation on activatable MMP‐2‐specific probes for optical imaging

Lebel, Réjean; Bonin, Marc-André; Zriba, Riadh; Radulska, Adrianna; Neugebauer, Witold; Lepage, Martin

doi:10.1002/cmmi.500

Cited by 5 publications

(11 citation statements)

References 28 publications

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“…Given the high concentration and size of albumin, the authors suggested that the probe would be largely bound to albumin and this interaction would therefore define the kinetics of their probe by causing unspecific accumulation through the EPR effect. We also reported on how albumin binding by some fluorescent probes may affect their optical properties . Hence, special care should be taken to assess unspecific protein binding of novel probes and its effect on accumulation and signal.…”

Section: Controls In Molecular Imagingmentioning

confidence: 99%

“…. Our group also recently published a set of Cy5/QSY21 MMP‐2 targeting agents . These smaller molecules are likely to penetrate more easily into tissues, but will be eliminated more quickly than most FF probes.…”

Section: Imaging Modalities and Probe Designsmentioning

confidence: 99%

“…Dynamic imaging of probes control molecules can provide information on P , K IN and K OUT . Our group recently published the in vitro characterization and in vivo stability of a series of signal‐modulated MMP‐2 specific fluorescent probes named I , II and III , along with their negative controls [ Is (scrambled), IId and IIId ( d ‐aa)] and a positive ( V ) control . Further in vivo results in a subcutaneous MC7‐L1 tumors in Balb/c mice were published separately .…”

Section: Controls In Molecular Imagingmentioning

confidence: 99%

“…Unlike V , IIId accumulates slowly over the first 50 min, and is then eliminated – which is perfectly in line with its more hydrophobic nature. This is caused by the quencher, QSY 21, which actually causes probes to aggregate in aqueous media. The behavior of this control is qualitatively similar to that of I , II , and III .…”

Section: Controls In Molecular Imagingmentioning

confidence: 99%

“…It is of course not possible to test in vitro against every macromolecule or enzyme of the host. Ex vivo plasma stability assays will provide a first‐hand evaluation of the in vivo stability of a probe . The two major limitations to this assay are the possibility that (a) the unspecific activation/degradation can occur in situ by membrane enzymes (i.e.…”

Section: Controls In Molecular Imagingmentioning

confidence: 99%

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A comprehensive review on controls in molecular imaging: lessons from MMP‐2 imaging

Lebel

Lepage

2014

Contrast Media & Molecular

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Metalloproteinases (MMPs), including MMP-2, play critical roles in tissue remodeling and are involved in a large array of pathologies, including cancer, arthritis and atherosclerosis. Their prognostic value warranted a large investment or resources in the development of noninvasive detection methods, based on probes for many current clinical and pre-clinical imaging modalities. However, the potential of imaging techniques is only matched by the complexity of the data they generate. This complexity must be properly assessed and accounted for in the early steps of probe design and testing in order to accurately determine the efficacy and efficiency of an imaging strategy. This review proposes basic rules for the evaluation of novel probes by addressing the specific case of MMP targeted probes.

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Section: Controls In Molecular Imagingmentioning

confidence: 99%

Section: Imaging Modalities and Probe Designsmentioning

confidence: 99%

Section: Controls In Molecular Imagingmentioning

confidence: 99%

Section: Controls In Molecular Imagingmentioning

confidence: 99%

Section: Controls In Molecular Imagingmentioning

confidence: 99%

See 3 more Smart Citations

A comprehensive review on controls in molecular imaging: lessons from MMP‐2 imaging

Lebel

Lepage

2014

Contrast Media & Molecular

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Deciphering Design Principles of Förster Resonance Energy Transfer-Based Protease Substrates: Thermolysin-Like Protease from Geobacillus stearothermophilus as a Test Case

et al. 2018

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Protease activity is frequently assayed using short peptides that are equipped with a Förster resonance energy transfer (FRET) reporter system. Many frequently used donor–acceptor pairs are excited in the ultraviolet range and suffer from low extinction coefficients and quantum yields, limiting their usefulness in applications where a high sensitivity is required. A large number of alternative chromophores are available that are excited in the visible range, for example, based on xanthene or cyanine core structures. These alternatives are not only larger in size but also more hydrophobic. Here, we show that the hydrophobicity of these chromophores not only affects the solubility of the resulting FRET-labeled peptides but also their kinetic parameters in a model enzymatic reaction. In detail, we have compared two series of 4–8 amino acid long peptides, designed to serve as substrates for the thermolysin-like protease (TLP-ste) from Geobacillus stearothermophilus . These peptides were equipped with a carboxyfluorescein donor and either Cy5 or its sulfonated derivative Alexa Fluor 647 as the acceptor. We show that the turnover rate k cat is largely unaffected by the choice of the acceptor fluorophore, whereas the K M value is significantly lower for the Cy5- than for the Alexa Fluor 647-labeled substrates. TLP-ste is a rather nonspecific protease with a large number of hydrophobic amino acids surrounding the catalytic site, so that the fluorophore itself may form additional interactions with the enzyme. This hypothesis is supported by the result that the difference between Cy5- and Alexa Fluor 647-labeled substrates becomes less pronounced with increasing peptide length, that is, when the fluorophore is positioned at a larger distance from the catalytic site. These results suggest that fluorophores may become an integral part of FRET-labeled peptide substrates and that K M and k cat values are generally only valid for a specific combination of the peptide sequence and FRET pair.

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Optimal dye-quencher pairs for the design of an “activatable” nanoprobe for optical imaging

Simard

Tomanek

Veggel

et al. 2013

Photochem Photobiol Sci

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Optical imaging offers high sensitivity and portability at low cost. The design of an optimal "activatable" imaging agent could greatly decrease the background noise and increase specificity of the signal. Five different molecules have been used to quench basal fluorescence of an enzyme substrate labeled with Cy5, Cy5.5 or IR800 at a distance of 8 amino acids (32 Å): a 6 nm gold nanoparticle (NP), a 20 nm and a 30 nm iron oxide (FeO) NP, the black hole quencher BHQ-3 and the IRdye quencher QC-1. The quenching efficiencies were 99% for QC1-IR800, 98% for QC1-Cy5.5, 96% for 30 nm FeO NP-Cy5.5, 89% for BHQ3-Cy5, 84% for BHQ3-Cy5.5, 77-90% for 6 nm gold NP-Cy5.5, depending on the number of dyes around the NP, 79% for 20 nm FeO NP-Cy5.5 and 77% for Cy5.5-Cy5. Signal activation upon cleavage by the matrix metalloproteinase MMP9 was proportional to the quenching efficiencies, ranging from 3-fold with Cy5.5-Cy5 to 67-fold with QC1-IR800. This independent work reports on the properties of the dyes and quenchers explaining the superior performance of QC-1 and 30 nm FeO NPs.

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Impact of H‐aggregation on activatable MMP‐2‐specific probes for optical imaging

Cited by 5 publications

References 28 publications

A comprehensive review on controls in molecular imaging: lessons from MMP‐2 imaging

A comprehensive review on controls in molecular imaging: lessons from MMP‐2 imaging

Deciphering Design Principles of Förster Resonance Energy Transfer-Based Protease Substrates: Thermolysin-Like Protease from Geobacillus stearothermophilus as a Test Case

Optimal dye-quencher pairs for the design of an “activatable” nanoprobe for optical imaging

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