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The assay quantifies glycolysis metabolism as an immune-cell activation biomarker, generated after antigenic stimulation with lung tumor–associated antigens. Results from two clinical trials show that an improved protocol increases diagnostic performance. 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Respiratory Research (2023) 24:52 [https://doi.org/10.1186/s12931-023-02358-w](https://doi.org/10.1186/s12931-023-02358-w)  \nRespiratory Research  \n RESEARCH Open Access  \nA novel, accurate, and non-invasive liquid  \nbiopsy test to measure cellular immune responses as a tool to diagnose early-stage lung cancer: a clinical trials study  \nShafrira Shai1*, Fernando Patolsky2, Hagai Drori 1, Eyal J. Scheinman 1, Eyal Davidovits 1, Giora Davidovits 1, Shoval Tirman1, Nadir Arber3, Amit Katz4 andYochai Adir5  \nAbstract  \nIntroduction Lung cancer remains the leading cause of death from cancer, worldwide. Developing early detection diagnostic methods, especially non-invasive methods, is a critical component to raising the overall survival rate and prognosis for lung cancer. The purpose of this study is to evaluate two protocols of a novel in vitro cellular immune response test to detect lung cancer. The test specifically quantifies the glycolysis metabolism pathway, which is abiomarker for the activation level of immune cells. It summarizes the results of two clinical trials, where each deploysa different protocol’s version of this test for the detection of lung cancer. In the later clinical trial, an improved test protocol is applied.  \nMethod The test platform is based on changes in the metabolic pathways of the immune cells following their activation by antigenic stimuli associated with Lung cancer. Peripheral Blood Mononuclear Cells are loaded on a multiwell plate together with various lung tumor associated antigens and a fluorescent probe that exhibits a pH-dependent absorption shift. The acidification process in the extracellular fluid is monitored by a commercial fluorescence plate reader device in continuous reading for 3 h at 37 °C to document the fluorescent signal received from each well. Results In the later clinical trial, an improved test protocol was applied and resulted in increased test accuracy. Specificity of the test increased to 94. 0% and test sensitivity increased to 97. 3% in lung cancer stage I, by using the improved protocol.  \nConclusion The improved protocol of the novel cellular immune metabolic response based test detects stage I and stage II of lung cancer with high specificity and sensitivity, with low material costs and fast results.  \nKeywords Early stage, Biomarker, Liquid biopsy, Tumor associated antigens, Immune cells metabolism  \n*Correspondence: Shafrira Shai[shafis@savicell.com](shafis@savicell.com)  \n1 Savicell Diagnostics Ltd., Matam Advanced Technology Park, Building  \n\\#23, P. O. Box 15050, 3190501 Haifa, Israel  \n2 School of Chemistry, Faculty of Exact Sciences, Tel Aviv University, 69978 Tel Aviv, Israel  \n3 Integrated Cancer Prevention Center, Tel Aviv Sourasky Medical Center, 6 Weizmann St., 6423906 Tel Aviv, Israel  \n4 Department of General Thoracic Surgery, Rambam Health Care Campus, 8 HaAliya HaShniya St., PO Box 9602, 31096 Haifa, Israel  \n5 Pulmonary Division, Faculty of Medicine, Lady Davis Carmel Medical Center, Mikhal Street 7, 3436212 Haifa, Israel  \n© The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/licenses/by/4.0/](http://creativecommons.org/licenses/by/4.0/. The Creative Co","cbCaikNbW2QlC96N","https://ap.wps.com/l/cbCaikNbW2QlC96N","pdf",2049135,11,"English","# Abstract\n# Introduction\n## Background and unmet need\n# Method\n## Test platform and measurement approach\n# Results\n## Performance of improved protocol\n# Conclusion\n# Keywords","[{\"question\":\"What is the purpose of the liquid biopsy test in this study?\",\"answer\":\"To evaluate two protocols of a novel in vitro cellular immune response test for detecting lung cancer, aiming at early-stage diagnosis using a non-invasive approach.\"},{\"question\":\"How does the test measure immune response?\",\"answer\":\"It quantifies glycolysis metabolism in immune cells after activation by lung tumor–associated antigens, using a pH-dependent fluorescent probe monitored continuously with a fluorescence plate reader.\"},{\"question\":\"What improvement was observed with the later clinical trial protocol?\",\"answer\":\"The improved protocol increased test accuracy, raising specificity to 94.0% and sensitivity to 97.3% for lung cancer stage I.\"}]","A novel, accurate, and non-invasive liquid biopsy test to measure cellular immune responses as a tool to diagnose early-stage lung cancer - a clinical trials study | PDF",28]