University of Angers, Inserm, CNRS, MINT, SFR ICAT, F-49000, Angers, France.
University of Rennes, CNRS, Inserm, BIOSIT, UMS 3480, US_S 018, F-35000, Rennes, France.
Pharm Res. 2021 Apr;38(4):681-691. doi: 10.1007/s11095-021-03034-8. Epub 2021 Apr 7.
Current preclinical therapeutic strategies involving nanomedicine require increasingly sophisticated nanosystems and the characterization of the complexity of such nanoassemblies is becoming a major issue. Accurate characterization is often the factor that can accelerate the translational approaches of nanomedicines and their pharmaceutical development to reach the clinic faster. We conducted a case study involving the adsorption of the NFL-TBS.40-63 (NFL) peptide (derived from neurofilaments) to the surface of lipid nanocapsules (LNCs) (a combined nanosystem used to target glioblastoma cells) to develop an analytical approach combining the separation and the quantification in a single step, leading to the characterization of the proportion of free peptide and thus the proportion of peptide adsorbed to the lipid nanocapsule surface.
LNC suspensions, NFL peptide solution and LNC/NFL peptide mixtures were characterized using a Size-Exclusion Chromatography method (with a chromatographic apparatus). In addition, this method was compared to centrifugal-filtration devices, currently used in literature for this case study.
Combining the steps for separation and characterization in one single sequence improved the accuracy and robustness of the data and led to reproducible results. Moreover the data deviation observed for the centrifugal-filtration devices demonstrated the limits for this increasingly used characterization approach, explained by the poor separation quality and highlighting the importance for the method optimization. The high potential of the technique was shown, proving that H-bond and/or electrostatic interactions mediate adsorption of the NFL peptide to the surface of LNCs.
Used only as a characterization tool, the process using chromatographic apparatus is less time and solvent consuming than classical Size-Exclusion Chromatography columns only used for separation. It could be a promising tool for the scientific community for characterizing the interactions of other combinations of nanosystems and active biological agents.
当前涉及纳米医学的临床前治疗策略需要越来越复杂的纳米系统，而此类纳米组装体的特性分析正成为一个主要问题。准确的特性分析通常是加速纳米药物转化方法及其药物开发、更快进入临床的关键因素。我们进行了一项案例研究，涉及 NFL-TBS.40-63（NFL）肽（源自神经丝）吸附到脂质纳米胶囊（LNC）表面的情况（一种用于靶向神经胶质瘤细胞的联合纳米系统），开发了一种分析方法，将分离和定量结合在单个步骤中，从而可以对游离肽的比例（因此也是吸附到脂质纳米胶囊表面的肽的比例）进行特性分析。
使用尺寸排阻色谱法（带有色谱仪）对 LNC 悬浮液、NFL 肽溶液和 LNC/NFL 肽混合物进行了表征。此外，还将该方法与目前文献中用于此案例研究的离心过滤装置进行了比较。
将分离和特性分析步骤组合在单个序列中提高了数据的准确性和稳健性，从而获得了可重复的结果。此外，离心过滤装置的数据偏差表明了该方法的局限性，这一局限性解释了这种日益广泛使用的特性分析方法的分离质量较差，并强调了方法优化的重要性。该技术的高潜力得到了证明，证明了氢键和/或静电相互作用介导了 NFL 肽对 LNC 表面的吸附。
仅作为特性分析工具，该方法使用色谱仪，比仅用于分离的传统尺寸排阻色谱柱更节省时间和溶剂。对于科学界来说，它可能是一种很有前途的工具，可用于对其他纳米系统和活性生物制剂组合的相互作用进行特性分析。
