Detection of activated platelets in a mouse model of carotid artery thrombosis with 18F-labeled single-chain antibodies

Ardipradja, Katie; Yeoh, Shinn Dee; Alt, Karen; O’Keefe, Graeme; Rigopoulos, Angela; Howells, David W.; Scott, Andrew M.; Peter, Karlheinz; Ackerman, Uwe; Hagemeyer, Christoph E.

doi:10.1016/j.nucmedbio.2013.12.006

Cited by 22 publications

(32 citation statements)

References 46 publications

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“…In addition, this approach is versatile. We have previously conjugated scFv with probes used in other imaging modalities and showed that thrombosis can be detected via targeted MRI 15, 16, ultrasound 14, 23 and PET 12 imaging. This advantage is of particular relevance in clinical settings, as these imaging facilities are available in more and more hospitals.…”

Section: Discussionmentioning

confidence: 99%

“…It merely provides information on the degree of luminal obstruction, but fails to inform the characteristics and stability of atherosclerotic plaques. We propose that targeting activated-platelets is an excellent strategy in the diagnosis of these diseases, and we have demonstrated their in vivo detection in an experimental mouse model of thrombosis using Positron Emission Tomography (PET) 12.…”

Section: Introductionmentioning

confidence: 99%

“…Consequently, it provides a better signal to background noise ratio and better access to target protein/cells. We have previously employed this scFv in various molecular imaging modalities such as intravital microscopy 13, ultrasound 14, PET 12 and MRI 15-18 of small animals. In these in viv o studies, no inhibitions of platelet aggregation, or bleeding prolongation or complications were observed with the dose used of the anti-activated-GPIIb/IIIa scFv.…”

Section: Introductionmentioning

confidence: 99%

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A Unique Recombinant Fluoroprobe Targeting Activated Platelets Allows In Vivo Detection of Arterial Thrombosis and Pulmonary Embolism Using a Novel Three-Dimensional Fluorescence Emission Computed Tomography (FLECT) Technology

Lim¹,

Yu²,

Huang³

et al. 2017

Theranostics

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Progress in pharmaceutical development is highly-dependent on preclinical in vivo animal studies. Small animal imaging is invaluable for the identification of new disease markers and the evaluation of drug efficacy. Here, we report for the first time the use of a three-dimensional fluorescence bioimager called FLuorescence Emission Computed Tomography (FLECT) for the detection of a novel recombinant fluoroprobe that is safe, easily prepared on a large scale and stably stored prior to scan. This novel fluoroprobe (Targ-Cy7) comprises a single-chain antibody-fragment (scFvTarg), which binds exclusively to activated-platelets, conjugated to a near-infrared (NIR) dye, Cy7, for detection. Upon mouse carotid artery injury, the injected fluoroprobe circulates and binds within the platelet-rich thrombus. This specific in vivo binding of the fluoroprobe to the thrombus, compared to its non-targeting control-fluoroprobe, is detected by the FLECT imager. The analyzed FLECT image quantifies the NIR signal and localizes it to the site of vascular injury. The detected fluorescence is further verified using a two-dimensional IVIS® Lumina scanner, where significant NIR fluorescence is detected in vivo at the thrombotic site, and ex vivo, at the injured carotid artery. Furthermore, fluorescence levels in various organs have also been quantified for biodistribution, with the highest fluoroprobe uptake shown to be in the injured artery. Subsequently, this live animal imaging technique is successfully employed to monitor the response of the induced thrombus to treatment over time. This demonstrates the potential of using longitudinal FLECT scanning to examine the efficacy of candidate drugs in preclinical settings. Besides intravascular thrombosis, we have shown that this non-invasive FLECT-imaging can also detect in vivo pulmonary embolism. Overall, this report describes a novel fluorescence-based preclinical imaging modality that uses an easy-to-prepare and non-radioactive recombinant fluoroprobe. This represents a unique tool to study mechanisms of thromboembolic diseases and it will strongly facilitate the in vivo testing of antithrombotic drugs. Furthermore, the non-radiation nature, low-cost, high sensitivity, and the rapid advancement of optical scanning technologies make this fluorescence imaging an attractive development for future clinical applications.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Unique Recombinant Fluoroprobe Targeting Activated Platelets Allows In Vivo Detection of Arterial Thrombosis and Pulmonary Embolism Using a Novel Three-Dimensional Fluorescence Emission Computed Tomography (FLECT) Technology

Lim¹,

Yu²,

Huang³

et al. 2017

Theranostics

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“…[7] In bacteria, Srt A cleaves an amide bond between Thr and Gly in the small pentapeptide recognition motif Leu-Pro-X-Thr-Gly (where X = any amino acid). [13] Herein, we describe the synthesis of new sarcophagine derivatives designed to function as substrates for Srt A and demonstrate their conjugation to scFv anti-LIBS , which is catalyzed by Srt A. The structure and function of antibodies is not compromised by the presence of the small sorting tag, and loss of biological activity is rarely observed.…”

mentioning

confidence: 99%

Enzyme‐Mediated Site‐Specific Bioconjugation of Metal Complexes to Proteins: Sortase‐Mediated Coupling of Copper‐64 to a Single‐Chain Antibody

et al. 2014

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The enzyme-mediated site-specific bioconjugation of a radioactive metal complex to a single-chain antibody using the transpeptidase sortase A is reported. Cage amine sarcophagine ligands that were designed to function as substrates for the sortase A mediated bioconjugation to antibodies were synthesized and enzymatically conjugated to a single-chain variable fragment. The antibody fragment scFv(anti-LIBS) targets ligand-induced binding sites (LIBS) on the glycoprotein receptor GPIIb/IIIa, which is present on activated platelets. The immunoconjugates were radiolabeled with the positron-emitting isotope (64)Cu. The new radiolabeled conjugates were shown to bind selectively to activated platelets. The diagnostic potential of the most promising conjugate was demonstrated in an in vivo model of carotid artery thrombosis using positron emission tomography. This approach gives homogeneous products through site-specific enzyme-mediated conjugation and should be broadly applicable to other metal complexes and proteins.

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“…The EGFR-targeted mAbs cetuximab and panitumumab were approved by the US Food and Drug Administration in 2004 and 2006, respectively, for the treatment of patients with metastatic colorectal cancer [5][6][7]. These include antigenbinding fragment (Fab; ∼ 50 kDa), single-chain antibody fragment (scFv; ∼ 30 kDa), scFv-CH3 dimer (minibody; ∼ 80 kDa), and single-domain antibody (sdAb; ∼ 15 kDa), also known as Nanobody [10][11][12][13]. However, because of the high molecular weight, the penetration of mAb into tumor tissue is poor.…”

Section: Introductionmentioning

confidence: 99%

99mTc-labeled single-domain antibody EG2 in targeting epidermal growth factor receptor

Wen

Wang

et al. 2015

Nuclear Medicine Communications

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Detection of activated platelets in a mouse model of carotid artery thrombosis with 18F-labeled single-chain antibodies

Cited by 22 publications

References 46 publications

A Unique Recombinant Fluoroprobe Targeting Activated Platelets Allows In Vivo Detection of Arterial Thrombosis and Pulmonary Embolism Using a Novel Three-Dimensional Fluorescence Emission Computed Tomography (FLECT) Technology

A Unique Recombinant Fluoroprobe Targeting Activated Platelets Allows In Vivo Detection of Arterial Thrombosis and Pulmonary Embolism Using a Novel Three-Dimensional Fluorescence Emission Computed Tomography (FLECT) Technology

Enzyme‐Mediated Site‐Specific Bioconjugation of Metal Complexes to Proteins: Sortase‐Mediated Coupling of Copper‐64 to a Single‐Chain Antibody

99mTc-labeled single-domain antibody EG2 in targeting epidermal growth factor receptor

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