Albumin-Binding Domain Conjugate for Near-Infrared Fluorescence Lymphatic Imaging
Albumin-Binding Domain Conjugate for Near-Infrared Fluorescence Lymphatic Imaging
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DOI：
10.1007/s11307-011-0499-x
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发表时间：
2012-06-01
影响因子：
3.1
通讯作者：
Sevick-Muraca, Eva M.
中科院分区：
文献类型：
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作者：
Davies-Venn, Cynthia A.;Angermiller, Bonnie;Sevick-Muraca, Eva M.
本研究的目的是开发和表征一种新型肽显像剂，用于无创近红外荧光成像淋巴中蛋白质运输。一种由环状白蛋白结合域（cABD）肽组成的显像剂，序列为Arg-Leu-Ile-Glu-Asp-Ile-Cys-Leu-Pro-Arg-Trp-Gly-Cys-Leu-Trp-Glu-Asp-Asp-Lys，与近红外荧光团IRDye800CW结合，增强血管摄取、保留和荧光成像。对cabd - irdye - 800肽偶联物进行了荧光光谱表征，以评估其光学性质，并使用SDS-PAGE和Biacore结合测定其结合亲和力和特异性。采用荧光成像法监测正常C57BL/6小鼠淋巴吸收和淋巴滞留情况。cABD-IRDye800比吲哚菁绿（一种以前用于人体淋巴血管成像的试剂）显示出大约6倍的荧光产量和更高的稳定性。与IRDye800CW相比，该制剂在纳米摩尔范围内对白蛋白具有IC50和Kd的亲和力，并且在小鼠淋巴中具有更好的保留特性。cABD-IRDye800可用于评估人类淋巴疾病小鼠模型中的淋巴功能，并有可能用于淋巴血管的临床诊断成像。
The aim of this study was to develop and characterize a novel peptide imaging agent for noninvasive near-infrared fluorescence imaging of protein transport by the lymphatics. An imaging agent consisting of a cyclic albumin-binding domain (cABD) peptide, with sequence, Arg-Leu-Ile-Glu-Asp-Ile-Cys-Leu-Pro-Arg-Trp-Gly-Cys-Leu-Trp-Glu-Asp-Asp-Lys, was conjugated to a near-infrared fluorophore, IRDye800CW, allowing for enhanced vascular uptake, retention, and fluorescence imaging.Characterization of the cABD-IRDye800 peptide conjugate was performed using fluorescence spectroscopy to assess optical properties and SDS-PAGE and Biacore binding assays to determine binding affinity and specificity. Fluorescence imaging of normal C57BL/6 mice was conducted to monitor lymphatic uptake and retention.cABD-IRDye800 exhibited approximately six times greater fluorescent yield and greater stability than indocyanine green, an agent previously used in humans to image lymphatic vasculature. The agent exhibited affinity for albumin with IC50 and Kd in the nanomolar range and demonstrated superior retention characteristics within mouse lymphatics when compared with IRDye800CW.cABD-IRDye800 has utility for assessing lymphatic function in mouse models of human lymphatic disease and the potential for use in clinical diagnostic imaging of the lymphatic vasculature.