Enhanced LSPR sensitivity for OpenSPR detection of therapeutic antibodies and polymer antibody conjugates

治疗性抗体代表了一类对癌症免疫治疗特别重要的药物。由于生物运输障碍，许多治疗由于缺乏到达疾病部位的抗体而失败。此外，通常施用大剂量的抗体，这可能导致毒副作用。因此，我们正在开发一种可注射的局部释放系统，以增加疾病部位的抗体浓度，同时降低副作用和成本。为了实现这些目标，我们需要快速检测和评估从我们的递送系统释放的治疗性抗体的活性的方法。Nicoya Lifesciences开发了第一个基于局部表面等离子体共振（LSPR）生物传感的商业仪器OpenSPR。与其他技术相比，OpenSPR具有高信噪比，适合于成本有效的高通量分析。目前，OpenSPR缺乏检测微微皮科至毫微微摩尔量抗体的灵敏度，而这是生物相关释放研究所需的。为此，我们将开发程序和材料，以增强OpenSPR仪器中抗体的LSPR检测。因为LSPR测量由于抗体结合到金OpenSPR表面后折射率变化引起的光吸收的偏移，所以LSPR增强将通过以下方式实现：（1）用吸收可见光的发色团修饰抗体;和（2）添加用吸收可见光的发色团修饰的抗体结合肽。Nicoya的LSPR技术与Wylie实验室在治疗性抗体方面的专业知识相结合，将导致新的LSPR方案和材料的开发，以快速分析稀释的抗体溶液。

Therapeutic antibodies represent a growing class of drug that are particularly important for cancer immunotherapies. Due to biological transport barriers, many treatments fail due to the lack of antibodies reaching the disease site. Furthermore, large doses of antibodies are commonly administered, which may lead to toxic side effects. Therefore, we are developing an injectable local release system to increase antibody concentrations at the disease site, while decreasing side effects and cost. To accomplish these goals, we need methods to rapidly detect and assess the activity of therapeutic antibodies released from our delivery systems. Nicoya Lifesciences has developed the first commercial instrument, OpenSPR, based on localized surface plasmon resonance (LSPR) biosensing. Compared to other techniques, OpenSPR has a high signal to noise ratios that is amenable to cost effective high throughput analysis. Currently, OpenSPR lacks the sensitivity to detect pico to femtomole amounts of antibodies, which is required for biologically relevant release studies. To this end, we will develop procedures and materials to enhance the LSPR detection of antibodies in the OpenSPR instrument. Because LSPR measures shifts in light absorption due to refractive index changes upon antibody binding to the gold OpenSPR surface, LSPR enhancement will be accomplished by: (1) modifying antibodies with chromophores that absorb visible light; and (2) adding antibody binding peptides modified with chromophores that absorb visible light. The combination of Nicoya's LSPR technology and the Wylie lab's expertise in therapeutic antibodies will result in the development of new LSPR protocols and materials to rapidly analyze dilute antibody solutions.****