RIKILT, Wageningen University & Research. P.O Box 230, 6700 AE Wageningen, The Netherlands.
Wageningen Food & Biobased Research, BioSensing & Diagnostics, Wageningen University & Research, P.O Box 17, 6700 AA, Wageningen, The Netherlands.
Biosensors (Basel). 2018 Dec 14;8(4):130. doi: 10.3390/bios8040130.
Lateral Flow Immunoassays (LFIAs) allow for rapid, low-cost, screening of many biomolecules such as food allergens. Despite being classified as rapid tests, many LFIAs take 10⁻20 min to complete. For a really high-speed LFIA, it is necessary to assess antibody association kinetics. By using a label-free optical technique such as Surface Plasmon Resonance (SPR), it is possible to screen crude monoclonal antibody (mAb) preparations for their association rates against a target. Herein, we describe an SPR-based method for screening and selecting crude anti-hazelnut antibodies based on their relative association rates, cross reactivity and sandwich pairing capabilities, for subsequent application in a rapid ligand binding assay. Thanks to the SPR selection process, only the fast mAb (F-50-6B12) and the slow (S-50-5H9) mAb needed purification for labelling with carbon nanoparticles to exploit high-speed LFIA prototypes. The kinetics observed in SPR were reflected in LFIA, with the test line appearing within 30 s, almost two times faster when F-50-6B12 was used, compared with S-50-5H9. Additionally, the LFIAs have demonstrated their future applicability to real life samples by detecting hazelnut in the sub-ppm range in a cookie matrix. Finally, these LFIAs not only provide a qualitative result when read visually, but also generate semi-quantitative data when exploiting freely downloadable smartphone apps.
侧向流动免疫分析（LFIAs）允许快速、低成本地筛选许多生物分子，如食物过敏原。尽管被归类为快速测试，但许多 LFIAs 需要 10-20 分钟才能完成。对于真正的高速 LFIA，有必要评估抗体结合动力学。通过使用无标记的光学技术，如表面等离子体共振（SPR），可以筛选粗制单克隆抗体（mAb）制剂，以评估它们与目标的结合速率。本文描述了一种基于 SPR 的方法，用于筛选和选择基于相对结合速率、交叉反应性和夹心配对能力的粗制抗榛子抗体，用于随后在快速配体结合测定中的应用。由于 SPR 选择过程，只有快速 mAb（F-50-6B12）和慢速 mAb（S-50-5H9）需要纯化，以便用碳纳米粒子进行标记，以利用高速 LFIA 原型。在 SPR 中观察到的动力学在 LFIA 中得到了反映，测试线在 30 秒内出现，当使用 F-50-6B12 时，比 S-50-5H9 快了近两倍。此外，这些 LFIA 还通过在曲奇基质中检测到亚 ppm 级别的榛子，证明了它们在现实生活样本中的未来应用前景。最后，这些 LFIA 不仅可以通过肉眼观察提供定性结果，还可以通过利用免费下载的智能手机应用程序生成半定量数据。
