Kota Krishna P, Trakht Ilya, Kalantarov Gavreel, Ordonez David, Wei Jiayi, Trefry Stephanie, Bavari Evia, Richardson Jenny, Zamani Rouzbeh, Raney Christy, Nasar Farooq, Daugherty Bruce, Lederman Seth, Bavari Sina
Tonix Pharmaceuticals, Frederick, MD 21701, USA.
Department of Medicine, Columbia University Medical Center, New York, NY 10032, USA.
Microorganisms. 2024 Jul 23;12(8):1503. doi: 10.3390/microorganisms12081503.
Convalescent sera, rich in pathogen-specific antibodies, offers passive immunity to patients with infectious diseases. Screening assays using convalescent sera are crucial for evaluating therapeutic efficacy, selecting suitable serum donors, and standardizing assays. They measure antibody levels, neutralizing potential, and specificity against viruses like SARS-CoV-2, ensuring therapeutic serum contains potent antibodies. Standardized procedures enable reliable results and wider adoption of serum therapy for COVID-19. We have developed a high-content image-based assay for screening convalescent sera against SARS-CoV-2 variants. Using various cell lines, we identified optimal candidates, employed immunofluorescence to visualize infected cells, and assessed neutralizing antibody efficacy. Screening convalescent sera for therapeutic potential identified neutralizing activity against SARS-CoV-2 variants. Dose-response analysis showed variable neutralizing activity, with some sera exhibiting broad neutralization. Additionally, we explored the synergy between neutralizing sera and β-d-N4-hydroxycytidine (NHC), an initial metabolite of molnupiravir. These assays enhance serum therapy's benefits for COVID-19 treatment and aid in understanding neutralizing activity against SARS-CoV-2 variants, addressing viral challenges.
恢复期血清富含病原体特异性抗体，可为传染病患者提供被动免疫。使用恢复期血清的筛选检测对于评估治疗效果、选择合适的血清供体以及规范检测至关重要。这些检测可测量抗体水平、中和潜力以及针对如SARS-CoV-2等病毒的特异性，确保治疗性血清含有强效抗体。标准化程序能够得出可靠结果，并使血清疗法在治疗COVID-19方面得到更广泛应用。我们开发了一种基于高内涵图像的检测方法，用于筛选针对SARS-CoV-2变体的恢复期血清。通过使用各种细胞系，我们确定了最佳候选血清，采用免疫荧光法观察感染细胞，并评估中和抗体的效力。对恢复期血清的治疗潜力进行筛选，确定了其对SARS-CoV-2变体的中和活性。剂量反应分析显示中和活性存在差异，一些血清表现出广泛的中和作用。此外，我们还探索了中和血清与β-d-N4-羟基胞苷（NHC，莫努匹拉韦的初始代谢产物）之间的协同作用。这些检测增强了血清疗法对COVID-19治疗的益处，并有助于了解针对SARS-CoV-2变体的中和活性，应对病毒挑战。
