High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease
| العنوان: | High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease |
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| المؤلفون: | Andrew C. Harris, Mark Vander Lugt, Aurelie Gomez, Praechompoo Pongtornpipat, Kelly Lamiman, Bryan Fiema, Sophie Paczesny |
| المصدر: | Journal of Visualized Experiments : JoVE Scopus-Elsevier |
| بيانات النشر: | MyJove Corporation, 2012. |
| سنة النشر: | 2012 |
| مصطلحات موضوعية: | General Chemical Engineering, medicine.medical_treatment, Small sample, Graft vs Host Disease, Enzyme-Linked Immunosorbent Assay, Pancreatitis-Associated Proteins, Disease, Hematopoietic stem cell transplantation, Blood plasma, Biology, Proteomics, Bioinformatics, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, proteomics, 0302 clinical medicine, Antigens, Neoplasm, Quantification, Acute graft versus host disease, Biomarkers, Tumor, graft-versus-host disease, medicine, Humans, Lectins, C-Type, Cytokine, 030304 developmental biology, 0303 health sciences, Issue 68, General Immunology and Microbiology, Plasma samples, Hepatocyte Growth Factor, General Neuroscience, Interleukin-2 Receptor alpha Subunit, biomarkers, medicine.disease, Elafin, High-Throughput Screening Assays, 3. Good health, surgical procedures, operative, Graft-versus-host disease, Sequential ELISA, Receptors, Tumor Necrosis Factor, Type I, Immunology, Medicine, ELISA, Complication, 030215 immunology |
| الوصف: | Unbiased discovery proteomics strategies have the potential to identify large numbers of novel biomarkers that can improve diagnostic and prognostic testing in a clinical setting and may help guide therapeutic interventions. When large numbers of candidate proteins are identified, it may be difficult to validate candidate biomarkers in a timely and efficient fashion from patient plasma samples that are event-driven, of finite volume and irreplaceable, such as at the onset of acute graft-versus-host disease (GVHD), a potentially life-threatening complication of allogeneic hematopoietic stem cell transplantation (HSCT). Here we describe the process of performing commercially available ELISAs for six validated GVHD proteins: IL-2Rα(5), TNFR1(6), HGF(7), IL-8(8), elafin(2), and REG3α(3) (also known as PAP1) in a sequential fashion to minimize freeze-thaw cycles, thawed plasma time and plasma usage. For this procedure we perform the ELISAs in sequential order as determined by sample dilution factor as established in our laboratory using manufacturer ELISA kits and protocols with minor adjustments to facilitate optimal sequential ELISA performance. The resulting plasma biomarker concentrations can then be compiled and analyzed for significant findings within a patient cohort. While these biomarkers are currently for research purposes only, their incorporation into clinical care is currently being investigated in clinical trials. This technique can be applied to perform ELISAs for multiple proteins/cytokines of interest on the same sample(s) provided the samples do not need to be mixed with other reagents. If ELISA kits do not come with pre-coated plates, 96-well half-well plates or 384-well plates can be used to further minimize use of samples/reagents. |
| تدمد: | 1940-087X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ae76459b8ab8b4d760d63fec9f8aca4f https://doi.org/10.3791/4247 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....ae76459b8ab8b4d760d63fec9f8aca4f |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1940087X |
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