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Dye-Loaded Polymersome-Based Lateral Flow Assay: Rational Design of a COVID-19 Testing Platform by Repurposing SARS-CoV-2 Antibody Cocktail and Antigens Obtained from Positive Human Samples
ACS Sensors ( IF 8.2 ) Pub Date : 2021-07-16 , DOI: 10.1021/acssensors.1c00854 Faezeh Ghorbanizamani 1 , Kerem Tok 1 , Hichem Moulahoum 1 , Duygu Harmanci 1 , Simge Balaban Hanoglu 1 , Ceren Durmus 1 , Figen Zihnioglu 1 , Serap Evran 1 , Candan Cicek 2 , Ruchan Sertoz 2 , Bilgin Arda 3 , Tuncay Goksel 4, 5 , Kutsal Turhan 6 , Suna Timur 1, 7
ACS Sensors ( IF 8.2 ) Pub Date : 2021-07-16 , DOI: 10.1021/acssensors.1c00854 Faezeh Ghorbanizamani 1 , Kerem Tok 1 , Hichem Moulahoum 1 , Duygu Harmanci 1 , Simge Balaban Hanoglu 1 , Ceren Durmus 1 , Figen Zihnioglu 1 , Serap Evran 1 , Candan Cicek 2 , Ruchan Sertoz 2 , Bilgin Arda 3 , Tuncay Goksel 4, 5 , Kutsal Turhan 6 , Suna Timur 1, 7
Affiliation
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The global pandemic of COVID-19 continues to be an important threat, especially with the fast transmission rate observed after the discovery of novel mutations. In this perspective, prompt diagnosis requires massive economical and human resources to mitigate the disease. The current study proposes a rational design of a colorimetric lateral flow immunoassay (LFA) based on the repurposing of human samples to produce COVID-19-specific antigens and antibodies in combination with a novel dye-loaded polymersome for naked-eye detection. A group of 121 human samples (61 serums and 60 nasal swabs) were obtained and analyzed by RT-PCR and ELISA. Pooled samples were used to purify antibodies using affinity chromatography, while antigens were purified via magnetic nanoparticles-based affinity. The purified proteins were confirmed for their specificity to COVID-19 via commercial LFA, ELISA, and electrochemical tests in addition to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. Polymersomes were prepared using methoxy polyethylene glycol-b-polycaprolactone (mPEG-b-PCL) diblock copolymers and loaded with a Coomassie Blue dye. The polymersomes were then functionalized with the purified antibodies and applied for the preparation of two types of LFA (antigen test and antibody test). Overall, the proposed diagnostic tests demonstrated 93 and 92.2% sensitivity for antigen and antibody tests, respectively. The repeatability (92–94%) and reproducibility (96–98%) of the tests highlight the potential of the proposed LFA. The LFA test was also analyzed for stability, and after 4 weeks, 91–97% correct diagnosis was observed. The current LFA platform is a valuable assay that has great economical and analytical potential for widespread applications.
中文翻译：
基于染料负载聚合物的侧向层析检测：通过重新利用从阳性人体样本中获得的 SARS-CoV-2 抗体混合物和抗原来合理设计 COVID-19 检测平台
COVID-19 的全球大流行仍然是一个重要威胁，特别是在发现新突变后观察到的快速传播率。从这个角度来看，及时诊断需要大量的经济和人力资源来减轻疾病。目前的研究提出了一种比色侧流免疫分析 (LFA) 的合理设计，其基础是重新利用人类样本来生产 COVID-19 特异性抗原和抗体，并结合用于肉眼检测的新型染料负载聚合物囊泡。获得一组 121 份人类样本（61 份血清和 60 份鼻拭子）并通过 RT-PCR 和 ELISA 进行分析。合并的样品用于使用亲和层析纯化抗体，而抗原则通过基于磁性纳米颗粒的亲和力纯化。除了十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析之外，还通过商业 LFA、ELISA 和电化学测试证实了纯化的蛋白质对 COVID-19 的特异性。使用甲氧基聚乙二醇-b-聚己内酯（mPEG- b -PCL）二嵌段共聚物制备聚合物囊泡，并负载考马斯蓝染料。然后用纯化的抗体对聚合物囊泡进行功能化，并用于制备两种类型的 LFA（抗原测试和抗体测试）。总体而言，所提出的诊断测试对抗原和抗体测试的敏感性分别为 93% 和 92.2%。测试的重复性 (92-94%) 和再现性 (96-98%) 凸显了所提出的 LFA 的潜力。还分析了 LFA 测试的稳定性，4 周后，观察到 91-97% 的正确诊断率。目前的 LFA 平台是一种有价值的检测方法，具有广泛应用的巨大经济和分析潜力。
更新日期：2021-08-27
中文翻译：
基于染料负载聚合物的侧向层析检测：通过重新利用从阳性人体样本中获得的 SARS-CoV-2 抗体混合物和抗原来合理设计 COVID-19 检测平台
COVID-19 的全球大流行仍然是一个重要威胁，特别是在发现新突变后观察到的快速传播率。从这个角度来看，及时诊断需要大量的经济和人力资源来减轻疾病。目前的研究提出了一种比色侧流免疫分析 (LFA) 的合理设计，其基础是重新利用人类样本来生产 COVID-19 特异性抗原和抗体，并结合用于肉眼检测的新型染料负载聚合物囊泡。获得一组 121 份人类样本（61 份血清和 60 份鼻拭子）并通过 RT-PCR 和 ELISA 进行分析。合并的样品用于使用亲和层析纯化抗体，而抗原则通过基于磁性纳米颗粒的亲和力纯化。除了十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析之外，还通过商业 LFA、ELISA 和电化学测试证实了纯化的蛋白质对 COVID-19 的特异性。使用甲氧基聚乙二醇-b-聚己内酯（mPEG- b -PCL）二嵌段共聚物制备聚合物囊泡，并负载考马斯蓝染料。然后用纯化的抗体对聚合物囊泡进行功能化，并用于制备两种类型的 LFA（抗原测试和抗体测试）。总体而言，所提出的诊断测试对抗原和抗体测试的敏感性分别为 93% 和 92.2%。测试的重复性 (92-94%) 和再现性 (96-98%) 凸显了所提出的 LFA 的潜力。还分析了 LFA 测试的稳定性，4 周后，观察到 91-97% 的正确诊断率。目前的 LFA 平台是一种有价值的检测方法，具有广泛应用的巨大经济和分析潜力。
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