The Taussson Ze team developed the SARS-CoV-2 protein chip and used it in serum IgG/IgM analysis in new crown patients.

The new epidemic is sweeping the world. Novel coronavirus pneumonia has become a global pandemic (pandemic) on 11 March,3, WHO (WHO) announced.detection of antibodies against new coronavirus, especially IgM, is helpful for rapid diagnosis of new coronavirus infection and is a powerful supplement to nucleic acid detection.at present, the identified antigens are mainly N and S proteins, and the antigenicity identification of other proteins of new coronavirus is still blank.in addition, exploring whether the antibody response intensity in patients is related to age or gender, disease progression and severity will help us to further understand the interaction between virus and immune system, which is of great significance for intervention therapy or vaccine development.novel coronavirus pneumonia patients are therefore required to systematically analyze the types and intensity of virus specific antibodies in the serum of patients with new coronavirus pneumonia.on March 27, 2020, Tao shengce team of Shanghai Jiaotong University and Zhou Jie team of Foshan Fourth People's Hospital jointly released the latest research results of global profiling of sars-cov-2 specific IgG / IgM responses of convalescent using a protein on medrxiv Microarray (based on sars-cov-2 protein chip, the global analysis of IgG / IgM antibody response in convalescent sera of new crown patients was performed).the team has synthesized a complete set of prokaryotic expression clones of sars-cov-2 (in order to make it easy for more scientists to obtain, the team has put this set of clones on the plasmid sharing platform of Suzhou Research Institute, Institute of biochemical cells, Chinese Academy of Sciences), and authorized them to share them later.the project team of Tao shengce of Shanghai Jiaotong University provided the prokaryotic expression clone of full set of coding gene of SARS CoV 2 for free). Novel coronavirus protein chip was successfully constructed byand based on the expressed clone library, the first SARS-CoV-2 protein chip was successfully constructed by Tao Shengce team.the chip currently covers the proteins encoded by 18 new coronavirus genomes, and many proteins have multiple versions and multiple sources, such as E.coli Expression, mammalian cell expression and cell-free expression.the chip layout, quality control and preliminary serum test results are shown in Figure 1.Figure 1. Layout of sars-cov-2 protein chip and serum analysis.A, chip layout and anti-6xhis quality control chart, in which the suffix in the protein name indicates the source of protein (T: Tao Lab); B: Hangzhou bioeast biotechnology. Co., Ltd.; K: healthcode Co., Ltd.; s: Sanyou biopharmaceuticals Co., Ltd.; W: Vacuum l Biotechnology Co., Ltd. Y: Sino biological Co., Ltd.; B, Based on this chip, the team conducted a comprehensive analysis of IgG and IgM antibodies in the serum of patients with new crown disease. The first 29 novel coronavirus pneumonia patients inwere from the Fourth People's Hospital of Foshan, including 3 mild cases, 26 ordinary patients, and no severe or severe patients.serum was collected on the day of discharge.in addition, 11 healthy controls and 10 lung cancer control samples were included. based on the sars-cov-2 proteomics chip, IgG and IgM were detected in each serum. the response profile of IgG is shown in Figure 2. through the cluster analysis of the response signals of each sample for all antigens, it was found that the patient group, the control group and the blank group were well clustered into three groups. Among them, S1 Protein and N protein generally had strong antibody response in patients, while almost all of the control group were negative, which confirmed the effectiveness of these two antigens in immunodiagnosis. further analysis showed that in the control group, two serum samples had strong signal against N protein, indicating that N protein may be false positive. in addition, the quantitative analysis of antibody response of proteins from different sources showed that both prokaryotic expression, mammalian cell expression or cell-free expression system had good detection effect, but the background of 293T protein in the control group was lower, or meant better discrimination effect (see the original text). The IgG profiles of 29 covid-19 patients, 21 control sera and blank controls showed strong response to other proteins of NCV in addition to S1 Protein and N protein. especially for protein orf9b, 13 cases were positive for IgG in 29 cases, 3 cases for nsp5, and 1 case for Nsp10, nsp14 and nsp16 for IgG (see the original text). as soon as possible, a small number of samples have been analyzed, but these preliminary results suggest that in addition to N and S proteins, other new coronavirus proteins may also produce antibodies, suggesting that these proteins may have important physiological or pathological effects. Fig. 3 Correlation analysis of age, LDH and other clinical characteristics with S1 Protein IgG response intensity. Finally, the correlation analysis was conducted between the IgG response of S1 and N protein and the clinical information of patients (Fig. 3). Excluding the influence of the time of onset (the time from the first symptom to sample acquisition), it was found that the antibody response of S1 or N protein was significantly positively correlated with age, especially for female patients In addition, the antibody response was positively correlated with the level of lactate dehydrogenase (LDH), and negatively correlated with the percentage of lymphocyte (ly%), especially in women. The antibody levels of three patients with mild illness were all at low levels, indicating that the antibody response was closely related to the severity of the disease. these observations or hints suggest that for young people, the body may be able to cope with it more easily, while for the elderly, it needs to mobilize a stronger immune response, and the female response may be more sensitive than the male. it is worth noting that the current analysis is still limited. More different types of samples and more comprehensive clinical association analysis are needed to obtain more definite conclusions. the chip is in continuous update. the comprehensive serum analysis based on the chip can help us to reveal the immune response of different populations after new coronavirus infection as soon as possible. in addition, the chip can also be used in, but not limited to, the following aspects: 1. Serological analysis. Novel coronavirus induced viral specific antibody response and its dynamic changes will be comprehensively studied by , which will help us understand the immune response process of the body and find the dominant protein antigen of the virus. 2. vaccine evaluation. dynamic monitoring of antibody levels against various protein components in serum after vaccination, and the correlation analysis between them and defense capacity will help the screening and early evaluation of vaccines, and accelerate the development process of vaccines. 3. Study on virus host interaction. to study the interaction between host key proteins and virus proteins, so as to help reveal the key mechanisms of virus infection, replication and synthesis, and provide potential target proteins for drug development and research. the co first authors of this study are Dr. Jiang Hewei, Dr. Li Yang, Dr. Zhang Hainan, and Dr. Wang Wei, the Fourth People's Hospital of Foshan, and the corresponding authors are Zhou Jie, President of Foshan Fourth People's Hospital and Tao Shengqi researcher of Shanghai Jiaotong University. other participants included Dr. Meng Dong, Wuhan Institute of virus, Chinese Academy of Sciences, and Dr. Yang Xiao, Institute of Biophysics, Chinese Academy of Sciences. original link: