Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies.
Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies.
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DOI：
10.1016/j.jim.2021.113082
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发表时间：
2021-08
影响因子：
2.2
通讯作者：
Gasque P
中科院分区：
文献类型：
--
作者：
Frumence E;Lebeau G;Viranaicken W;Dobi A;Vagner D;Lalarizo Rakoto M;Sandenon Seteyen AL;Giry C;Septembre-Malaterre A;Raffray L;Gasque P
开发新的诊断检测方法成为管理COVID-19的优先事项。为此，我们在这里提出了一个内部ELISA的基础上生产的两个主要的重组和高质量的抗原从SARS-CoV-2。在CHO细胞系中产生与小鼠IgG 2a-Fc融合的全长N和S-RBD片段蛋白。分泌的重组蛋白易于用标准蛋白A色谱法纯化，并用于内部ELISA中，以检测COVID-19 RTPCR阳性患者血浆中的抗N和抗RBD IgG。在所有阳性血浆样品中容易检测到对重组抗原的高反应性，而对照健康受试者的血浆中未观察到识别。值得注意的是，从细胞培养上清液中获得的未纯化的重组N蛋白也适用于通过ELISA监测阳性患者中的IgG水平。本工作为研制低成本高质量的血清学试剂盒提供了初步的前景。
The development of new diagnostic assays become a priority for managing COVID-19. To this aim, we presented here an in-house ELISA based on the production of two major recombinant and high-quality antigens from SARS-CoV-2. Full-length N and S-RBD fragment proteins fused to mouse IgG2a-Fc were produced in the CHO cell line. Secreted recombinant proteins were easily purified with standard Protein A chromatography and were used in an in-house ELISA to detect anti-N and anti-RBD IgGs in the plasma of COVID-19 RTPCR-positive patients. High reactivity against recombinant antigens was readily detected in all positive plasma samples, whereas no recognition was observed with control healthy subject's plasmas. Remarkably, unpurified recombinant N protein obtained from cell culture supernatant was also suitable for the monitoring by ELISA of IgG levels in positive patients. This work provides an early prospection for low price but high-quality serological kit development.
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影响因子：
5.6
作者：
Diemer C;Schneider M;Seebach J;Quaas J;Frösner G;Schätzl HM;Gilch S
通讯作者：
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影响因子：
7.8
作者：
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通讯作者：
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DOI：
10.1016/s0006-291x(02)00566-1
发表时间：
2002-06-21
影响因子：
3.1
作者：
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影响因子：
64.5
作者：
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