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mINImIZING EXCITATION LIGHT LEAKAGE AND mAXImIZING mEASUREmENT SENSITIVITY FOR mOLECULAR ImAGING WITH NEAR-INFRARED FLUORESCENCE

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Journal of Innovative Optical Health Sciences 2011年第3期4卷 301-307页

作者： BANGHE ZHU eva m.sevick-muraca Center for molecular Imaging The Brown Foundation Institute of molecular medicine The University of Texas Health Science Center at Houston Houston TX 77030USA

Near-infraredfluorescence(NIRF)imaging involves the separation of weakfluorescence signals from backscattered excitation *** measurement sensitivity of current NIRF imaging systems is limited by the excitation light leakage through *** this contribution,the authors demonstrate that the excitation light leakage can be suppressed upon using appropriatefilter combination and *** excitation light leakage and measurement sensitivity were assessed and compared in this study by computing the transmission ratios of excitation to emission light collected and the signal-to-noise ratios in well-controlled phantom studies with differentfilter combinations and *** appropriatefilter combinations and permutations,we observe as much as two orders of magnitude reduction in the transmission ratio and higher signal-to-noise ratio.

关键词： molecular imaging fluorescence opticalfilter excitation light leakage noise floor

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Influence of chelator and near-infrared dye labeling on biocharacteristics of dual-labeled trastuzumab-based imaging agents

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Chinese Journal of Cancer Research 2016年第3期28卷 362-369页

作者： Xuejuan Wang melissa B Aldrich Zhi Yang Nina Zhou Qing Xie Chen Liu eva sevick-muraca Key Laboratory of Carcinogenesis and Translational Research(ministry of Education/Beijing) Department of Nuclear MedicinePeking University Cancer Hospital&InstituteBeijing 100142China Center for molecular Imaging the Brown Foundation Institute of Molecular MedicineUniversity of Texas Health Science Center at HoustonHouston TX 77030USA

Objective： To investigate the effect of fluorescent dye labeling on the targeting capabilities of 111 In- （DTPA）.-trastuzumab-（IRDye 800）m. methods： Trastuzumab-based conjugates were synthesized and conjugated with diethylenetriaminepentaacetic acid （DTPA） at molar ratios of I, 2, 3 and 5 and with a fluorescent dye （IRDye 800CW） at molar ratios of 1, 3 and 5. Immunoreactivity and internalization were assessed on SKBR-3 cells, overexpressing human epidermal growth factor receptor 2. The stability in human serum and phosphate-buffered saline （PBS） was evaluated. The biodistribution of dual-labeled conjugates was compared with that of 111In-（DTPA）2-trastuzumab in a SKBR-3 xenograft model to evaluate the effect of dye-to-protein ratio. Results： All trastuzumab-based conjugates exhibited a high level of chemical and optical purity. Flow cytometry results showed that increasing dye-to-protein ratios were associated with decreased immunoreactivity. Stability studies revealed that the conjugate was stable in PBS, while in human serum, increased degradation and protein precipitation were observed with increasing dye-to-protein ratios. At 4 h, the percentages of internalization of dual-labeled conjugates normalized by dye-to-protein ratio （m） were 24.88%±2.10%, 19.99%±0.59%, and 17.47%±1.26% for ＂m＂ equal to 1, 3, and 5, respectively. A biodistribution study revealed a progressive decrease in tumor uptake with an increase in the dye-to-protein ratios. The liver, spleen and kidney showed a marked uptake with increased dye-to-protein ratios, particularly in the latter. Conclusions： With non-specific-site conjugation of the fluorescent dye with a protein based on imaging agent, the increase in dye-to-protein ratios negatively impacted the immunoreactivity and stability, indicating a reduced tumor uptake.

关键词： Influence near infrared dye trastuzumab human epidermal growth factor receptor 2 dual-labeled

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Preclinical characterization and validation of a dual-labeled trastuzumab-based imaging agent for diagnosing breast cancer

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Chinese Journal of Cancer Research 2015年第1期27卷 74-82页

作者： Xuejuan Wang melissa B. Aldrich milton V. marshall eva m. sevick-muraca Department of Nuclear medicine Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education) Peking University Cancer Hospital & Institute Division of molecular Imaging The Brown Foundation Institute of Molecular Medicine University of Texas Health Science Center at Houston

Objective： The combination of both nuclear and fluorescent reporters provides unique opportunities for noninvasive nuclear imaging with subsequent fluorescence image-guided resection and pathology. Our objective was to synthesize and optimize a dual-labeled trastuzumab-based imaging agent that can be used to validate an optical imaging agent with potential use in identifying tumor metastases in human epidermal growth factor receptor 2 （HER2） positive breast cancer patients. methods： [111In]-DTPA-trastuzumab-IRDye 800 was synthesized by a three-step procedure. Purity, stability, immunoreactivity, internalization and biodistribution were explored in HER2＋ SKBR-3 cells. Biodistribution of [111In]-DTPA-trastuzumab-IRDye 800 was performed in a SKBR-3 xenograft model. Results： [111In]-DTPA-trastuzumab-IRDye 800 demonstrated high purity by both chemical and fluorometric determinations. Both flow cytometry and the Lindmo assay demonstrated a high binding affinity of [111In]-DTPA-trastuzumab-IRDye 800 to HER2-overexpressing cells. The dual-labeled conjugate was stable in PBS, but not in serum after 24 h at 37 ℃. Larger molecules （〉150 kD） were seen after a 24 h-incubation in human serum. Biodistribution studies revealed tumor-specific accumulation of [111In]-DTPA- trastuzumab-IRDye 800 in SKBR-3 tumors, and tumor uptakes at 24 and 48 h were （12.42±1.72）% and （9.96±1.05） %, respectively, following intravenous administration. The tumor-to-muscle ratio was 9.13±1.68 at 24 h, and increased to 12.79±2.13 at 48 h. Liver and kidney showed marked uptake of the dual-labeled imaging agent. Conclusions： [111In]-DTPA-trastuzumab-IRDye 800 is an effective diagnostic biomarker that can be used to validate dual-labeled, molecularly targeted imaging agents and can allow these agents to be translated into clinical practice for identifying HER2＋ lesions.

关键词： Dual-labeling trastuzumab human epidermal growth factor receptor 2 (HER2) near-infrared opticalimaging (NIR optical imaging) radioimmunoscintigraphy

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