Journal of immunological methods最新文献_第10页

Novel hapten design, highly sensitive monoclonal antibody production, and immunoassay development for rapid screening of illegally added chloramphenicol in cosmetics 用于快速筛查化妆品中非法添加的氯霉素的新型合体设计、高灵敏度单克隆抗体的生产和免疫测定法的开发

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-22 DOI: 10.1016/j.jim.2023.113604

Zhaoxiang Wang , Mian Wang , Xiaoxiang Fu , Jingqi Qian , Min Wang , Guiyu Tan

{"title":"Novel hapten design, highly sensitive monoclonal antibody production, and immunoassay development for rapid screening of illegally added chloramphenicol in cosmetics","authors":"Zhaoxiang Wang , Mian Wang , Xiaoxiang Fu , Jingqi Qian , Min Wang , Guiyu Tan","doi":"10.1016/j.jim.2023.113604","DOIUrl":"10.1016/j.jim.2023.113604","url":null,"abstract":"<div><p><span><span>Hapten design and synthesis have been regarded as the key factor to generate high-quality antibodies. In the present study, a novel hapten of chloramphenicol was synthesized, characterized and compared with two conventional haptens. The new hapten generated mAb 4B5 showed higher sensitivity and titer than the other two haptens-based mAbs. The haptens synthesized with the structure of chloramphenicol base generated more sensitive antibodies than the hapten with chloramphenicol succinate, and the spacer arm linked to the phenyl group hapten elicited the strongest </span>antibody response. After optimization, a direct competitive enzyme-linked immunosorbent assay (dcELISA) and a lateral flow </span>immunoassay<span> (LFIA), both based on the mAb 4B5, were developed. The dcELISA had a half maximum inhibition concentration<span> of 0.23 ng/mL and the LFIA showed a cutoff value of 5–10 ng/mL. The LFIA was applied to detect illegally-added chloramphenicol samples in anti-acne cosmetics, five out of 19 samples were tested chloramphenicol containing within 10 min, which result was confirmed with the dcELISA and HPLC. The LFIA has an adequate sensitivity and can be used as a point of care diagnostic device for rapidly screening chloramphenicol in cosmetics.</span></span></p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139024401","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Novel hapten design, highly sensitive monoclonal antibody production, and immunoassay development for rapid screening of illegally added chloramphenicol in cosmetics 用于快速筛查化妆品中非法添加的氯霉素的新型合体设计、高灵敏度单克隆抗体的生产和免疫测定法的开发

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-22 DOI: 10.1016/j.jim.2023.113604

Zhaoxiang Wang, Mian Wang, Xiaoxiang Fu, Jingqi Qian, Min Wang, Guiyu Tan

{"title":"Novel hapten design, highly sensitive monoclonal antibody production, and immunoassay development for rapid screening of illegally added chloramphenicol in cosmetics","authors":"Zhaoxiang Wang, Mian Wang, Xiaoxiang Fu, Jingqi Qian, Min Wang, Guiyu Tan","doi":"10.1016/j.jim.2023.113604","DOIUrl":"https://doi.org/10.1016/j.jim.2023.113604","url":null,"abstract":"<p>Hapten design and synthesis have been regarded as the key factor to generate high-quality antibodies. In the present study, a novel hapten of chloramphenicol was synthesized, characterized and compared with two conventional haptens. The new hapten generated mAb 4B5 showed higher sensitivity and titer than the other two haptens-based mAbs. The haptens synthesized with the structure of chloramphenicol base generated more sensitive antibodies than the hapten with chloramphenicol succinate, and the spacer arm linked to the phenyl group hapten elicited the strongest antibody response. After optimization, a direct competitive enzyme-linked immunosorbent assay (dcELISA) and a lateral flow immunoassay (LFIA), both based on the mAb 4B5, were developed. The dcELISA had a half maximum inhibition concentration of 0.23 ng/mL and the LFIA showed a cutoff value of 5–10 ng/mL. The LFIA was applied to detect illegally-added chloramphenicol samples in anti-acne cosmetics, five out of 19 samples were tested chloramphenicol containing within 10 min, which result was confirmed with the dcELISA and HPLC. The LFIA has an adequate sensitivity and can be used as a point of care diagnostic device for rapidly screening chloramphenicol in cosmetics.</p>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139022489","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Dissociation of murine oral mucosal tissues for single cell applications 解离小鼠口腔黏膜组织，用于单细胞应用

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-22 DOI: 10.1016/j.jim.2023.113605

Tomoko Ikeuchi , Ramin Akhi , Belmaliz Cardona Rodriguez , David Fraser , Drake Williams , Tae Sung Kim , Teresa Greenwell-Wild , Andrew Overmiller , Maria Morasso , Niki Moutsopoulos

{"title":"Dissociation of murine oral mucosal tissues for single cell applications","authors":"Tomoko Ikeuchi , Ramin Akhi , Belmaliz Cardona Rodriguez , David Fraser , Drake Williams , Tae Sung Kim , Teresa Greenwell-Wild , Andrew Overmiller , Maria Morasso , Niki Moutsopoulos","doi":"10.1016/j.jim.2023.113605","DOIUrl":"10.1016/j.jim.2023.113605","url":null,"abstract":"<div><p><span><span>Single-cell RNA sequencing and flow cytometry approaches have been instrumental in understanding cellular states within various tissues and organs. However, tissue dissociation methods can potentially alter results and create bias due to preferential recovery of particular cell types. Here we present efforts to optimize methods for dissociation of murine oral </span>mucosal tissues and provide three different protocols that can be utilized to isolate major cell populations in the </span>oral mucosa<span>. These methods can be used both in health and in states of inflammation, such as periodontitis<span>. The optimized protocols use different enzymatic approaches (collagenase II, collagenase IV and the Miltenyi whole skin dissociation kit) and yield preferential recovery of immune, stromal and epithelial cells, respectively. We suggest choosing the dissociation method based on the cell population of interest to study, while understanding the limitations of each approach.</span></span></p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139020054","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells 从猕猴个体记忆 B 细胞中分离单克隆抗体的 SMART 方法

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-14 DOI: 10.1016/j.jim.2023.113602

Jason T. Weinfurter , Sarah N. Bennett , Matthew R. Reynolds

{"title":"A SMART method for isolating monoclonal antibodies from individual rhesus macaque memory B cells","authors":"Jason T. Weinfurter , Sarah N. Bennett , Matthew R. Reynolds","doi":"10.1016/j.jim.2023.113602","DOIUrl":"10.1016/j.jim.2023.113602","url":null,"abstract":"<div><p>Characterizing antigen-specific B cells is a critical component of vaccine and infectious disease studies in rhesus macaques (RMs). However, it is challenging to capture immunoglobulin variable (IgV) genes from individual RM B cells using 5′ multiplex (MTPX) primers in nested PCR reactions. In particular, the diversity within RM IgV gene leader sequences necessitates large 5′ MTPX primer sets to amplify IgV genes, decreasing PCR efficiency. To address this problem, we developed a switching mechanism at the 5′ ends of the RNA transcript (SMART)-based method for amplifying IgV genes from single RM B cells to capture Ig heavy and light chain pairs. We demonstrate this technique by isolating simian immunodeficiency virus (SIV) envelope-specific antibodies from single-sorted RM memory B cells. This approach has several advantages over existing methods for cloning antibodies from RMs. First, optimized PCR conditions and SMART 5′ and 3′ rapid amplification of cDNA ends (RACE) reactions generate full-length cDNAs from individual B cells. Second, it appends synthetic primer binding sites to the 5′ and 3′ ends of cDNA during synthesis, allowing for PCR amplification of low-abundance antibody templates. Third, the nested PCR primer mixes are simplified by employing universal 5′ primers, eliminating the need for complex 5′ MTPX primer sets. We anticipate this method will enhance the isolation of antibodies from individual RM B cells, supporting the genetic and functional characterization of antigen-specific B cells.</p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0022175923001849/pdfft?md5=78cdbcda96db334a8d1db5dfbdf65ca7&pid=1-s2.0-S0022175923001849-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138685727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Variegate expression of Cre recombinase in hematopoietic cells in CD11c-cre transgenic mice CD11c-cre 转基因小鼠造血细胞中 Cre 重组酶的差异表达

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-14 DOI: 10.1016/j.jim.2023.113600

Claire Murat , Sylvie Guerder

引用次数: 0

Tumor nano-lysate activates dendritic cells to evoke a preventative immune response 肿瘤纳米溶液激活树突状细胞，唤起预防性免疫反应

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-12 DOI: 10.1016/j.jim.2023.113601

Jenna A. Dombroski , Abigail R. Fabiano , Samantha V. Knoblauch , Schyler J. Rowland , Katherine N. Gibson-Corley , Michael R. King

{"title":"Tumor nano-lysate activates dendritic cells to evoke a preventative immune response","authors":"Jenna A. Dombroski , Abigail R. Fabiano , Samantha V. Knoblauch , Schyler J. Rowland , Katherine N. Gibson-Corley , Michael R. King","doi":"10.1016/j.jim.2023.113601","DOIUrl":"10.1016/j.jim.2023.113601","url":null,"abstract":"<div><p>A tumor nano-lysate “TNL” vaccine comprised of sonicated 4T1 cells was developed, characterized and implemented for the prevention of triple-negative breast cancer. This study aimed to gain a better understanding of the immune response behind the success of the vaccine in vivo, through use of ex vivo and in vivo assays. Here, we analyze the activation of various immune cells isolated from healthy mouse spleens and find that antigen-presenting cells (APCs) such as dendritic cells (DCs) are being activated following 24 h incubation with 1:10 mg TNL/mg splenocytes. These cells were further explored to determine the pathway by which activation is occurring, and it was observed that TNL are phagocytosed by DCs to activate NF-kB and c-Fos pathways, resulting in enhanced cytokine release after 24 h. An in vivo temporal analysis was performed in mice to understand the immune response at 1, 3, 7 and 10 days after one 100 μL dose of TNL consisting of 10<sup>5</sup> sonicated 4T1 cells via cardiac puncture and splenocyte and peripheral blood mononuclear cell (PBMC) analysis. Changes were observed for up to one week. A multiple dose study was performed comparing mice that were vaccinated with one dose of TNL administered every ten days for 3 doses total, as well as a PBS vehicle control. Survival for TNL-vaccinated mice was enhanced compared to the PBS control, and there was an average delay of 10 days in the onset of metastasis. The differences between the groups at the end of the study demonstrate the potential for TNL as a preventative therapeutic.</p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0022175923001837/pdfft?md5=c9b8cc3df8e88cb0e17965abb6ab0856&pid=1-s2.0-S0022175923001837-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138577006","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Application of enzyme-linked immunosorbent assay to detect antimicrobial peptides in human intestinal lumen 应用酶联免疫吸附试验检测人体肠腔中的抗菌肽

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-09 DOI: 10.1016/j.jim.2023.113599

Julie S. Hong , Abrar Shamim , Hussein Atta , Eric B. Nonnecke , Sarah Merl , Satyajit Patwardhan , Elin Manell , Esad Gunes , Philip Jordache , Bryan Chen , Wuyuan Lu , Bo Shen , Beatrice Dionigi , Ravi P. Kiran , Megan Sykes , Emmanuel Zorn , Charles L. Bevins , Joshua Weiner

{"title":"Application of enzyme-linked immunosorbent assay to detect antimicrobial peptides in human intestinal lumen","authors":"Julie S. Hong , Abrar Shamim , Hussein Atta , Eric B. Nonnecke , Sarah Merl , Satyajit Patwardhan , Elin Manell , Esad Gunes , Philip Jordache , Bryan Chen , Wuyuan Lu , Bo Shen , Beatrice Dionigi , Ravi P. Kiran , Megan Sykes , Emmanuel Zorn , Charles L. Bevins , Joshua Weiner","doi":"10.1016/j.jim.2023.113599","DOIUrl":"10.1016/j.jim.2023.113599","url":null,"abstract":"<div><p><span><span>Intestinal transplantation is the definitive treatment for intestinal failure. However, tissue rejection and graft-versus-host disease are relatively common complications, necessitating aggressive immunosuppression that can itself pose further complications. Tracking intraluminal markers in ileal effluent from standard ileostomies may present a noninvasive and sensitive way to detect developing pathology within the intestinal graft. This would be an improvement compared to current assessments, which are limited by poor sensitivity and specificity, contributing to under or over-immunosuppression, respectively, and by the need for invasive biopsies. Herein, we report an approach to reproducibly analyze ileal fluid obtained through stoma sampling for antimicrobial peptide/protein concentrations, reasoning that these molecules may provide an assessment of intestinal </span>homeostasis<span> and levels of intestinal inflammation over time. Concentrations of lysozyme (LYZ), </span></span>myeloperoxidase (MPO), calprotectin (S100A8/A9) and β-defensin 2 (DEFB2) were assessed using adaptations of commercially available enzyme-linked immunosorbent assays (ELISAs). The concentration of α-defensin 5 (DEFA5) was assessed using a newly developed sandwich ELISA. Our data support that with proper preparation of ileal effluent specimens, precise and replicable determination of antimicrobial peptide/protein concentrations can be achieved for each of these target molecules via ELISA. This approach may prove to be reliable as a clinically useful assessment of intestinal homeostasis over time for patients with ileostomies.</p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138565788","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In silico epitope prediction of Borrelia burgdorferi sensu lato antigens for the detection of specific antibodies 为检测特异性抗体而对包柔氏病原抗原的表位进行硅学预测

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-07 DOI: 10.1016/j.jim.2023.113596

Weronika Grąźlewska, Karolina Sołowińska, Lucyna Holec-Gąsior

{"title":"In silico epitope prediction of Borrelia burgdorferi sensu lato antigens for the detection of specific antibodies","authors":"Weronika Grąźlewska, Karolina Sołowińska, Lucyna Holec-Gąsior","doi":"10.1016/j.jim.2023.113596","DOIUrl":"10.1016/j.jim.2023.113596","url":null,"abstract":"<div><p><span><span>Despite many years of research, serodiagnosis of Lyme disease still faces many obstacles. Difficulties arise mainly due to the low degree of </span>amino acid sequence conservation of the most immunogenic antigens among </span><em>B. burgdorferi</em><span> s.l. genospecies, as well as differences in protein production depending on the environment in which the spirochete<span> is located. Mapping B-cell epitopes located on antigens allows for a better understanding of antibody-pathogen interactions which is essential for the development of new and more effective diagnostic tools. In this study, in silico B-cell epitope mapping was performed to determine the theoretical diagnostic potential of selected </span></span><em>B. burgdorferi</em><span> s.l. proteins (BB0108, BB0126, BB0298, BB0689, BB0323, FliL, PstS, SecD, EF-Tu). Bioinformatics software<span> predicted 35 conserved linear and 31 conformational epitopes with the degree of identity among </span></span><em>B. burgdorferi</em> s.l. of at least 85%, which may prove to be useful in the development of a new tool for the diagnosis of Lyme disease.</p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138557093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evaluation of HLA-DR expression in monocytes and CD64 in neutrophils as A predictor of SEPSIS/sirs in the infectious-inflammatory process 评估单核细胞中HLA-DR表达和中性粒细胞中CD64表达作为感染-炎症过程中败血症/sirs的预测因子。

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-12-01 DOI: 10.1016/j.jim.2023.113589

Clara Giffoni de Freitas , Mariela Granero Farias

{"title":"Evaluation of HLA-DR expression in monocytes and CD64 in neutrophils as A predictor of SEPSIS/sirs in the infectious-inflammatory process","authors":"Clara Giffoni de Freitas , Mariela Granero Farias","doi":"10.1016/j.jim.2023.113589","DOIUrl":"10.1016/j.jim.2023.113589","url":null,"abstract":"<div><p>Sepsis is a highly fatal disease that affects millions of people worldwide every year. Currently, the diagnosis of sepsis is made by identifying at least two symptoms of systemic inflammatory response syndrome (SIRS), along with confirming the presence of microorganisms using a blood culture examination. Some biomarkers are already used to aid in the diagnosis, such as increased levels of C-reactive protein (CRP), leukocytes, immature granulocytes<span><span> (IG), and bands. In addition, studies have shown a relationship between the expression of certain antigen receptors in the body's defense cells and its infectious state. CD64 is a receptor expressed in monocytes, and, in cases of infection, its expression is strongly observed in </span>neutrophils. On the other hand, the class II MHC (major histocompatibility complex) marker, HLA-DR (human leukocyte antigen-DR), decreases its expression in monocytes in response to infection. This cohort study was conducted with 77 adult patients from a university hospital, divided into two groups: Non-Sepsis/SIRS and Sepsis/SIRS. The selected samples were analyzed by flow cytometry, identifying the expression of CD64 and HLA-DR according to their MFI, and calculating the sepsis index (SI) for each patient. All three parameters exhibited significant differences in expression between the two groups. When compared to the laboratory tests already in use, the utilization of HLA-DR, CD64, and the new index has shown greater sensitivity and specificity in identifying sepsis. This study contributes to knowledge about the relationship between the expression of antigens on defense cells and sepsis. The use of these biomarkers can help to improve the diagnosis and treatment of sepsis, which may contribute to the reduction of mortality related to the disease.</span></p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138477874","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of automated microfluidic immunoassays for the detection of SARS-CoV-2 antibodies and antigen 自动微流控免疫法检测SARS-CoV-2抗体和抗原的建立。

IF 2.2 4区医学

Journal of immunological methods Pub Date : 2023-11-30 DOI: 10.1016/j.jim.2023.113586

Linwood Johnson , Maggie L. Bartlett , Francisco Ramirez , Christopher D. Heger , Darci R. Smith

{"title":"Development of automated microfluidic immunoassays for the detection of SARS-CoV-2 antibodies and antigen","authors":"Linwood Johnson , Maggie L. Bartlett , Francisco Ramirez , Christopher D. Heger , Darci R. Smith","doi":"10.1016/j.jim.2023.113586","DOIUrl":"10.1016/j.jim.2023.113586","url":null,"abstract":"<div><p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the coronavirus disease 2019 (COVID-19) global pandemic. Rapid and sensitive detection of the virus soon after infection is important for the treatment and prevention of transmission of COVID-19, and detection of antibodies is important for epidemiology, assessment of vaccine immunogenicity, and identification of the natural reservoir and intermediate host(s). Patient nasal or oropharyngeal swabs or saliva used in conjunction with polymerase chain reaction (PCR) detect SARS-CoV-2 RNA, whereas lateral flow immunoassays (LFI) detect SARS-CoV-2 proteins. Enzyme-linked immunosorbent assays (ELISA) detect anti-SARS-CoV-2 antibodies in blood. Although effective, these assays have poor sensitivity (e.g., LFI) or are labor intensive and time consuming (PCR and ELISA). Here we describe the development of rapid, automated ELISA-based immunoassays to detect SARS-CoV-2 antigens and antibodies against the virus. The Simple Plex™ platform uses rapid microfluidic reaction kinetics for sensitive analyte detection with small sample volumes. We developed three sensitive <90-min Simple Plex immunoassays that measure either the SARS-CoV-2 antigens or the immune response to SARS-CoV-2, including neutralizing antibodies, in serum from COVID-19 patients.</p></div>","PeriodicalId":16000,"journal":{"name":"Journal of immunological methods","volume":null,"pages":null},"PeriodicalIF":2.2,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0022175923001680/pdfft?md5=74116bdadc8ca52bdd6bd54e07dbf294&pid=1-s2.0-S0022175923001680-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138470303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0