Octet® 非标记分子互作分析系统采用实时的非标记分析技术，用于测定亲和力、动力学和抗体/蛋白质定量。这种无流路的仪器平台基于生物层干涉（BLI）技术而设计，与传统的非标记技术相比，具有众多优势。

2024年上半年（截至6月30日在线发表）的国内论文中，使用赛多利斯Octet® 数据的国内论文呈现井喷状态。陈老师统计了2024年上半年约200篇国内论文（预计总数超过200篇），这些文章分布于130个单位， 详细列表在文末哦~老规矩，让我们先按照单位和应用进行统计。
 

图1. 2024上半年Octet® 文章第一作者单位分布
 

图2. 2024H1 Octet® 文章应用分布
 
中国药科大学、北京协和医学院、南京大学、复旦大学、浙江大学和中山大学等单位发表的文章均在4篇以上。其中，中国药科大学论文数达到了10篇。这表明，Octet® 因其操作简单、成本低、数据准确和仪器稳定性好等优点，可以帮助科学家快速取得科研成果。

从应用方向上看，依旧涉及到多个方向。其中小分子的应用超过蛋白和抗体应用的文章数量，牢牢占据应用方向榜首！BLI技术对溶剂（DMSO）不敏感，假阳性低，高灵敏度等优点可以帮助研究人更快，更高质量的获得了小分子检测的结果。


今天，我们就来挑选几篇代表性文章来一一解读：


浙江大学——细胞与分子互作^[1]
 
全球气候变化伴随着二氧化碳（CO₂）富集和高温（HT）胁迫。研究发现，番茄外源质外糖 （Glc）处理增强了番茄在环境CO₂条件下对HT胁迫的恢复力。基于细胞的生物层干涉法（Octet® ）、亚细胞定位和分裂荧光素酶测定表明，Glc 与番茄 G 蛋白信号转导 1 （RGS1） 调节因子结合并诱导 RGS1 内吞作用，从而以浓度依赖性方式诱导 RGS1-G 蛋白α亚基 （GPA1） 解离，这是 CO₂-Glc 诱导的耐热性升高所必需的。这些信息增强了对Glc-G蛋白信号传导功能在应对全球气候变化的胁迫恢复力中的理解，并将有助于开发植物恢复力的基因工程方法。
 

图3. 将原生质体固化在Octet® 传感器上，结合各种单糖，发现表达RGS1的细胞主要结合葡萄糖，但是将RGS1突变后，与葡萄糖结合变弱
 
Octet® 因为浸入即读的方法，更适合检测比较大的分析物，比较细胞。


西湖大学——血清抗体浓度测定^[2]
 
第三剂灭活新型冠状病毒疫苗的抗体反应对疫苗的保护作用至关重要。西湖大学工学院对接种疫苗的15人的九个月里七个时间点进行了微量取样和静脉穿刺取样，并建立了基于Octet® 的光纤生物层干涉法测量(FO-BLI)来测试血清中的中和抗体，可以在几分钟内完成测试。研究结果揭示发现在第三次给药后，野生型变体的血清NAb水平在7天后增加了2.9倍，在一个月内增加了3.3倍，随后下降，并在三个月后变得不可检测。针对原始株与变异株的中和能力检测, FO-BLI与假病毒中和试验高度相关(Pearson 相关系数为0.983)。基于FO-BLI的浓度测试因为其快速，并且与功能性匹配，有潜力在疾病预防和疫苗开发中有着更大的应用。
 

图4. A图：Octet® 建立血清中和抗体的检测方法，将受体ACE2结合在传感器上，与HRP偶联的不同病毒株的RBD和血清的混合物孵育，然后用底物放大信号；B图：这种方法测试的标准抗体的灵敏度可以达到20ng/mL；
 

D图：FO-BLI的检测结果与病毒中和实验PVNT（IC50）有良好的相关性

使用生物层干涉技术，可以快速建立浓度检测方法。
 
 
浙江工业大学/北大医学院——分子垂钓^[3]

研究者建立了一种基于Octet® 的垂钓技术，研究人员将GNAS蛋白固化至SA传感器表面，再与中药参芪降糖颗粒（SJG）提取物结合，随后提取物被解离至洗脱液中，将洗脱液使用UHPLC-Q/TOF-MS/MS分析GNAS垂钓出的小分子。通过垂钓组与对照组的相对含量变化筛选26个小分子可能与GNAS有相互作用。其中7个化合物用Octet® 进一步验证，并显示出强的结合活性，均是SJG的主要成分。其中，黄芪中的Formononetin、高丽参的三七皂苷Ft1和人参皂苷中的甲素人参F2为“君“、地黄中的Catapol为“臣”，五味子中的五味子素A和茜草中的没食子酸为“佐使”，符合中医配伍“君臣佐使“原则。
 

图5. Octet® 验证小分子和GNAS的结合
 
非流路设计可以多次垂钓富集丰度低结合弱的小分子，提高垂钓的成功率。


中国医学科学院基础医学研究所——脂肪肝新靶点^[4]
 
肝细胞对预防非酒精性脂肪性肝病（NAFLD）和其他慢性代谢性疾病具有重要意义。中国医学科学院基础医学研究所黄波团队研究发现，糖原合成使用其中间代谢物尿苷二磷酸葡萄糖（UDPG）来拮抗脂肪生成，从而引导小鼠和人肝细胞将葡萄糖碳储存为糖原。在这项机制研究中，发现UDPG通过SLC35F5转运进入高尔基体后，诱导site-1蛋白酶S1P的泛素化降解，从而阻断活性形式SREBP1c的生成，最终抑制脂肪酸的合成。通过Octet® 非标记分子互作系统对小鼠和人的脂肪酸合成过程中关键酶（S1P）与UDPG结合这一重要步骤进行了验证，并提供了结合的直观证据。
 

图6. 通过SA传感器固化小鼠（a）和人（b）的S1P蛋白分别与不同浓度UDPG结合解离曲线

原文大量pulldown以及CO-IP数据，但作者仍选择用Octet® 非标记互作系统来证明S1P与UDPG直接相互作用，可见Direct Binding是趋势。
 
 
Octet® 分子互作分析系统的优势在于

• 非标记Direct Binding是趋势，结果更准确
• 快速测定亲和力，更加定量化地表征分子互作
• 无洗涤步骤，可测弱亲和力（解离快）
• 写入了美国药典，文章多，认可度广
• 万金油技术，可以用与检测DNA，小分子，蛋白质等各种生物分子
• 多种实验方案，除了直接测试，还有竞争法，垂钓等
• 操作简便，耗材及维护成本低

Octet® 让分子互作不再复杂！祝愿大家可以用赛多利斯神器发好文章！
    
《生物层干涉技术小分子应用文集》详细介绍了生物层干涉（BLI）技术的原理，及其在小分子领域的多个应用场景，并对BLI技术应用于小分子研究的20余篇代表性文献进行了整理汇总：内容涵盖从防御机制到垂钓验证，再到小分子筛选、表征、优化等多个方面，全展示了BLI技术在小分子研究中的强大潜力和实践成果。
  
 
-参考文献-
[1] Glucose-G protein signaling plays a crucial role in tomato resilience to high temperature and elevated CO2 | Plant Physiology | Oxford Academic (oup.com)
[2] Vaccines | Free Full-Text | Dynamic Profiling and Prediction of Antibody Response to SARS-CoV-2 Booster-Inactivated Vaccines by Microsample-Driven Biosensor and Machine Learning (mdpi.com)
[3] Zhang H, et al. Discovery of drug targets based on traditional Chinese medicine microspheres (TCM-MPs) fishing strategy combined with bio-layer interferometry (BLI) technology. Anal Chim Acta. 2024 May 29;1305:342542.
[4] Hepatic glycogenesis antagonizes lipogenesis by blocking S1P via UDPG.Science,2024

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https://www.sciencedirect.com/science/article/abs/pii/S0043135424008091
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https://www.biorxiv.org/content/10.1101/2024.06.04.597356v1.full.pdf
https://www.mdpi.com/2076-2607/12/6/1148
https://www.sciencedirect.com/science/article/pii/S0753332224007108
https://www.pnas.org/doi/abs/10.1073/pnas.2400163121
https://onlinelibrary.wiley.com/doi/abs/10.1002/ange.202406677
https://www.sciencedirect.com/science/article/pii/S2590157524004176
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https://www.nature.com/articles/s41420-024-02005-3
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https://www.sciencedirect.com/science/article/abs/pii/S0026265X24009585
https://www.cell.com/cell-reports/fulltext/S2211-1247(24)00593-X
https://www.sciencedirect.com/science/article/abs/pii/S0039914024006258
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