Barreto Kris, Bernhard Wendy, Toledo Darien, Jett Kimberly, Casaco Angel, León Kalet, Geyer C Ronald
Department of Pathology and Laboratory Medicine, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada.
Center of Molecular Immunology (CIM), Havana, Cuba.
Mol Imaging Biol. 2025 Jun 17. doi: 10.1007/s11307-025-02026-z.
Targeted and broadly applicable molecular targets are important for image guided surgery. Xenoantigens represent a particularly interesting class of targets. This study evaluates the xenoantigen N-glycolyl-neuraminic acid GM3 ganglioside (Neu5Gc-GM3) as a potential fluorescence-guided surgical tool.
The antibody 14F7hT is conjugated to the near-infrared dye (IRDye800CW) and characterized under GLP conditions. The quality and stability of the 14F7hT-IRDye800CW probe was assessed. In vivo imaging using 14F7hT-IRDye800CW in mice with Neu5Gc GM3 positive and negative xenografts were compared to a control IgG-IRDye800CW probe targeting an epitope not present on the xenografts. Biodistribution, pharmacokinetics, and toxicity were evaluated.
The 14F7hT-IRDye800CW probe was 98 ± 2% pure as determined by micro-capillary electrophoresis. The KDapp as determined by binding cell-lines expressing the target was unchanged after conjugation. We demonstrate a peak accumulation window of 12 - 48 h in murine xenografts with male and female CD-1 nude mice administered 0.5 nmoles of the probe (i.v.) and very low uptake in other tissues. Preclinical toxicity studies in male and female balb/c mice support a no observed adverse effect level (NOAEL) of 50 mg/kg in mice.
The 14F7hT-IRDye800CW probe was found to be safe and have low non-specific uptake in a model organism known to express the target. These data support future clinical development of the probe.
靶向且广泛适用的分子靶点对图像引导手术很重要。异种抗原是一类特别有趣的靶点。本研究评估异种抗原N-羟乙酰神经氨酸GM3神经节苷脂（Neu5Gc-GM3）作为一种潜在的荧光引导手术工具。
抗体14F7hT与近红外染料（IRDye800CW）偶联，并在GLP条件下进行表征。评估14F7hT-IRDye800CW探针的质量和稳定性。将14F7hT-IRDye800CW在具有Neu5Gc GM3阳性和阴性异种移植瘤的小鼠体内成像与靶向异种移植瘤上不存在的表位的对照IgG-IRDye800CW探针进行比较。评估生物分布、药代动力学和毒性。
通过微毛细管电泳测定，14F7hT-IRDye800CW探针的纯度为98±2%。偶联后，通过结合表达靶点的细胞系测定的KDapp未发生变化。我们证明，在给雄性和雌性CD-1裸鼠静脉注射0.5纳摩尔探针后，小鼠异种移植瘤中在12至48小时出现峰值蓄积窗口，而在其他组织中的摄取量非常低。对雄性和雌性balb/c小鼠进行的临床前毒性研究支持小鼠的无观察到不良反应水平（NOAEL）为50mg/kg。
在已知表达靶点的模型生物体中，发现14F7hT-IRDye800CW探针是安全的，且非特异性摄取较低。这些数据支持该探针未来的临床开发。
