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

治疗性抗体代表了一类日益增长的药物，对癌症免疫治疗尤其重要。由于生物运输障碍，许多治疗由于缺乏抗体到达疾病部位而失败。此外，通常使用大剂量的抗体，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，同时减少副作用和成本。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活性的方法。Nicoya生命科学公司开发了第一款基于局部表面等离子体共振（LSPR）生物传感的商用仪器OpenSPR。与其他技术相比，OpenSPR具有高信噪比，可用于经济高效的高吞吐量分析。目前，OpenSPR缺乏检测微到飞摩尔抗体量的灵敏度，而这是生物学相关释放研究所必需的。为此，我们将开发程序和材料来增强OpenSPR仪器中抗体的LSPR检测。由于LSPR测量的是抗体与金OpenSPR表面结合时折射率变化引起的光吸收变化，因此LSPR的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；(2)加入吸收可见光的发色团修饰的抗体结合肽。Nicoya的LSPR技术和Wylie实验室在治疗性抗体方面的专业知识的结合将导致新的LSPR方案和材料的开发，以快速分析稀释抗体溶液。****

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.****