Rapid and quantitative detection of SARS-CoV-2 specific IgG for convalescent serum evaluation
Rapid and quantitative detection of SARS-CoV-2 specific IgG for convalescent serum evaluation
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DOI：
10.1016/j.bios.2020.112572
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发表时间：
2020-12-01
影响因子：
12.6
通讯作者：
Fan, Xudong
中科院分区：
文献类型：
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作者：
Tan, Xiaotian;Krel, Mila;Fan, Xudong
具有高丰度中和IgG的恢复期血清是一种很有前途的治疗剂，用于抢救处于危急阶段的COVID-19患者。了解SARS-CoV-2 Sl特异性IgG的浓度对于选择合适的恢复期血清供体至关重要。在这里，我们提出了一种便携式微流控ELISA技术，用于快速（15分钟），定量和灵敏的检测抗SARS-CoV-2 S1 IgG在人血清中，只有8 μ L的样品体积。我们首先鉴定了一种人源化单克隆IgG，其对SARS-CoV-2 S1蛋白具有较高的结合亲和力和相对较高的特异性，其随后可作为抗SARS-CoV-2 S1 IgG的血清学分析的校准标准。然后，我们测量了16名恢复期COVID-19患者中抗SARS-CoV-2 S1 IgG的丰度。由于校准标准品的可用性和我们测定的大动态范围，我们能够仅用一个固定稀释因子（200 x）鉴定用于恢复期血清治疗的“合格供体”。最后，我们证明了我们的技术可以在40分钟内灵敏地检测SARS-CoV-2抗原（S1和N蛋白），灵敏度为pg/mL。总体而言，我们的技术可以大大促进快速，灵敏和定量分析COVID-19相关标志物，用于治疗，诊断，流行病学和预后目的。
Convalescent serum with a high abundance of neutralization IgG is a promising therapeutic agent for rescuing COVID-19 patients in the critical stage. Knowing the concentration of SARS-CoV-2 Sl-specific IgG is crucial in selecting appropriate convalescent serum donors. Here, we present a portable microfluidic ELISA technology for rapid (15 min), quantitative, and sensitive detection of anti-SARS-CoV-2 S1 IgG in human serum with only 8 mu L sample volume. We first identified a humanized monoclonal IgG that has a high binding affinity and a relatively high specificity towards SARS-CoV-2 S1 protein, which can subsequently serve as the calibration standard of anti-SARS-CoV-2 S1 IgG in serological analyses. We then measured the abundance of anti-SARS-CoV-2 S1 IgG in 16 convalescent COVID-19 patients. Due to the availability of the calibration standard and the large dynamic range of our assay, we were able to identify "qualified donors" for convalescent serum therapy with only one fixed dilution factor (200 x). Finally, we demonstrated that our technology can sensitively detect SARS-CoV-2 antigens (S1 and N proteins) with pg/mL level sensitivities in 40 min. Overall, our technology can greatly facilitate rapid, sensitive, and quantitative analysis of COVID-19 related markers for therapeutic, diagnostic, epidemiologic, and prognostic purposes.