{"id":2752,"date":"2024-05-27T14:47:01","date_gmt":"2024-05-27T06:47:01","guid":{"rendered":"https:\/\/opentrons.com.cn\/?post_type=knowledge2&p=2752"},"modified":"2024-09-03T17:47:36","modified_gmt":"2024-09-03T09:47:36","slug":"opentrons-elisa","status":"publish","type":"knowledge2","link":"https:\/\/en.opentrons.com.cn\/knowledge2\/opentrons-elisa\/","title":{"rendered":"Automated Elisa (enzyme-linked immunosorbent assay) using Opentrons pipetting workstations"},"content":{"rendered":"\n

Introduction<\/strong> ELISA (enzyme-linked immunosorbent assay) is an important test method widely used in diagnostics and research. In this technology, ensuring consistency in sample processing is critical, and reducing hands-on time can effectively reduce human error. We developed and tested multiple automation protocols using Opentrons OT-2 and Flex liquid handling workstations. These automated protocols allow you to perform experiments directly using ELISA kits, or you can customize specific experimental procedures according to your needs.<\/p>\n\n\n\n

Method<\/strong><\/p>\n\n\n\n

\"\u68c0\u6d4b\u65b9\u6cd5\u793a\u610f\u56fe\"<\/figure>\n\n\n\n

Kit: Takara Fibronectin (FN) EIA Kit (sandwich ELISA, Figure 1)<\/p>\n\n\n\n

    \n
  • Instrument: Opentrons OT-2<\/li>\n\n\n\n
  • Workflow:<\/li>\n<\/ul>\n\n\n\n
      \n
    1. Add sample (targeted capture)<\/li>\n\n\n\n
    2. washing<\/li>\n\n\n\n
    3. Add enzyme-labeled antibody<\/li>\n\n\n\n
    4. washing<\/li>\n\n\n\n
    5. Add chromogenic substrate<\/li>\n<\/ol>\n\n\n\n
      \n

      Detection method: Tecan Cortisol Saliva ELISA Kit (competition ELISA, Figure 2)<\/p>\n\n\n\n

      \"\u68c0\u6d4b\u65b9\u6cd5\u793a\u610f\u56fe\"<\/figure>\n\n\n\n
        \n
      • Instrument: Opentrons OT-2<\/li>\n\n\n\n
      • Workflow:<\/li>\n<\/ul>\n\n\n\n
          \n
        1. Add sample (target capture) & label antigen<\/li>\n\n\n\n
        2. washing<\/li>\n\n\n\n
        3. Add chromogenic substrate<\/li>\n<\/ol>\n<\/div>\n\n\n\n

          <\/p>\n\n\n\n

            \n
          • \u68c0\u6d4b\u65b9\u6cd5: Cell Sciences SARS-CoV-2 Surrogate Virus Neutralization Test Kit
            \uff08\u7ade\u4e89\u6cd5ELISA\uff0c\u56fe3\uff09<\/li>\n\n\n\n
          • Instrument: Opentrons OT-2<\/li>\n\n\n\n
          • Workflow:<\/li>\n<\/ul>\n\n\n\n
              \n
            1. Loading (Targeted Capture) & Labeling ACE2<\/li>\n\n\n\n
            2. washing<\/li>\n\n\n\n
            3. Add chromogenic substrate<\/li>\n<\/ol>\n\n\n\n
              \"\u68c0\u6d4b\u65b9\u6cd5\u793a\u610f\u56fe\"<\/figure>\n\n\n\n
                \n
              • \u68c0\u6d4b\u65b9\u6cd5: SARS-CoV-2 Spike RBD-coupled Magnetic Beads
                (\u78c1\u73e0\u5f0fELISA, \u56fe4)<\/li>\n\n\n\n
              • Instruments: Opentrons OT-2 & Magnetic Stand Module<\/li>\n\n\n\n
              • Workflow:<\/li>\n<\/ul>\n\n\n\n
                  \n
                1. Add sample (targeted capture)<\/li>\n\n\n\n
                2. washing<\/li>\n\n\n\n
                3. Add enzyme labeled antibody<\/li>\n\n\n\n
                4. washing<\/li>\n\n\n\n
                5. Add chromogenic substrate<\/li>\n<\/ol>\n\n\n\n
                  \"\u68c0\u6d4b\u65b9\u6cd5\u793a\u610f\u56fe\"<\/figure>\n\n\n\n
                  \"ELISA
                  Figure 5<\/figcaption><\/figure>\n\n\n\n

                  Detection: Assay method development for cytokine detection (sandwich ELISA, Figure 5) Instrument: Opentrons Flex Workflow: 1. ELISA plate preparation 2. Washing 3. Adding sample (target capture) 4. Washing 5. Adding enzyme-labeled antibody 6. Washing 7. Adding substrate to develop color<\/p>\n\n\n\n

                  Results<\/strong><\/p>\n\n\n\n

                  \"\u7ed3\u679c\u663e\u793a\u56fe\"<\/figure>\n\n\n\n

                  Figure 6. Takara Fibronectin (FN) EIA Kit (Figure 1). The FN sample was gradient diluted and tested. The test results are as shown in the figure: the left picture shows the FN sample concentration and average absorbance (n=3) data, showing a highly linear regression; the right picture shows the results of three repeated experiments, showing good repeatability. sex and stability.<\/p>\n\n\n\n

                  \"\u7ed3\u679c\u663e\u793a\u56fe\"<\/figure>\n\n\n\n

                  Figure 7. Tecan Cortisol Saliva ELISA Kit (Figure 2). The cortisol sample was gradient diluted and tested. The test results are as shown in the figure: The left picture shows the cortisol sample concentration and average absorbance (n=3) data, showing a highly linear regression; the right picture shows the results of three repeated experiments, which are well presented. repeatability and stability.<\/p>\n\n\n\n

                  \"\"<\/figure>\n\n\n\n

                  Figure 8. Cell Sciences SARS-CoV-2 Surrogate (Figure 3). The test results show that the positive human serum sample (the ELISA test result that has been previously certified by the FDA is positive) shows a positive result in this test (inhibition rate >20%, calculation formula: inhibition rate = [1 - (sample OD450 value\/OD value of negative control)] x 100%, n = 3).<\/p>\n\n\n\n

                  \"\"<\/figure>\n\n\n\n

                  Figure 9. SARS-CoV-2 Spike RBD-coupled Magnetic Beads (Figure 4). Recombinant neutralizing antibody samples were serially diluted and tested. Test results show that positive human serum samples, previously tested positive in an FDA-cleared ELISA assay, tested positive in this test (n = 3) (right).<\/p>\n\n\n\n

                  \"\"<\/figure>\n\n\n\n

                  Figure 10. Assay Development for Cytokine Detection (Figure 5). Anti-TNF-alpha antibodies and anti-IFN-gamma antibodies were coated on a 96-well plate, and samples containing cytokines were tested by ELISA. The data are as shown in the figure (n=6).<\/p>\n\n\n\n

                  Conclusion<\/strong><\/p>\n\n\n\n

                    \n
                  • This workstation can be used to perform the complete ELISA workflow from adding antibody\/antigen to adding chromogenic substrate<\/li>\n\n\n\n
                  • \u7ed3\u679c\u8868\u660e\u6837\u672c\u5904\u7406\u7684\u4e00\u81f4\u6027\u9ad8\uff08\u53d8\u5f02\u7cfb\u6570<10%\uff09\u3001\u51c6\u786e\u6027\u9ad8\uff08R-squared>0.99\uff09\uff0c\u91cd\u590d\u6027
                    \u597d\u3002<\/li>\n\n\n\n
                  • \u4ee5 SARS-CoV-2 \u6297\u4f53\u68c0\u6d4b\u4e3a\u4f8b\uff0c\u57fa\u4e8e 96 \u5b54\u677f\u7684\u5546\u7528\u8bd5\u5242\u548c\u57fa\u4e8e\u78c1\u73e0\u7684\u81ea\u5236 ELISA \u8bd5\u5242\u5747\u53ef
                    \u5b9e\u73b0\u5168\u81ea\u52a8\u5316\u5904\u7406\uff0c\u4e14\u7ed3\u679c\u4e0e FDA \u8ba4\u8bc1\u7684 Elisa \u65b9\u6cd5\u6d4b\u5b9a\u7ed3\u679c\u4e00\u81f4\u3002<\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"

                    Introduction ELISA (enzyme-linked immunosorbent assay) […]<\/p>\n","protected":false},"featured_media":2570,"template":"","meta":{"inline_featured_image":false,"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0},"knowledge2_cat":[96],"knowledge2_app":[],"class_list":["post-2752","knowledge2","type-knowledge2","status-publish","has-post-thumbnail","hentry","knowledge2_cat-biji"],"yoast_head":"\n\u4f7f\u7528 Opentrons \u79fb\u6db2\u5de5\u4f5c\u7ad9\u8fdb\u884c\u81ea\u52a8\u5316Elisa\uff08\u9176\u8054\u514d\u75ab\u5438\u9644\u8bd5\u9a8c\uff09 - Opentrons\u4e2d\u56fd\u5b98\u7f51<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/en.opentrons.com.cn\/knowledge2\/opentrons-elisa\/\" \/>\n<meta property=\"og:locale\" content=\"zh_CN\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"\u4f7f\u7528 Opentrons \u79fb\u6db2\u5de5\u4f5c\u7ad9\u8fdb\u884c\u81ea\u52a8\u5316Elisa\uff08\u9176\u8054\u514d\u75ab\u5438\u9644\u8bd5\u9a8c\uff09 - 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