دورية أكاديمية
A novel luciferase-based assay for the detection of Chimeric Antigen Receptors.
| العنوان: | A novel luciferase-based assay for the detection of Chimeric Antigen Receptors. |
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| المؤلفون: | Gopalakrishnan R; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Matta H; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Choi S; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Natarajan V; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Prins R; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Gong S; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Zenunovic A; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Narasappa N; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Patel F; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Prakash R; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Chaudhary V; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Sikri V; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Chitnis SD; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Kochegarov A; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Wang D; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Falat M; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Kahn M; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Keerthipati PS; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Sharma N; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Lenka J; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Stieben TM; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Braun J; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Batra A; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Purvis K; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Ito K; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Lee JH; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Jeronimo A; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Zamora HM; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Membreno A; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Qiu Q; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Peshin S; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Namburu L; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America., Chaudhary PM; Jane Anne Nohl Division of Hematology and Center for the Study of Blood Diseases, University of Southern California, Keck School of Medicine, Los Angeles, California, United States of America. preet.chaudhary@med.usc.edu. |
| المصدر: | Scientific reports [Sci Rep] 2019 Feb 13; Vol. 9 (1), pp. 1957. Date of Electronic Publication: 2019 Feb 13. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: London : Nature Publishing Group, copyright 2011- |
| مواضيع طبية MeSH: | Immunotherapy, Adoptive/*methods , Luciferases/*metabolism , Receptors, Chimeric Antigen/*analysis, Cell Line ; Flow Cytometry/methods ; Humans ; Lymphocytes/metabolism ; Receptors, Antigen/metabolism ; Receptors, Antigen, T-Cell/metabolism ; Receptors, Chimeric Antigen/metabolism ; Recombinant Fusion Proteins/metabolism ; T-Lymphocytes/immunology |
| مستخلص: | Chimeric Antigen Receptor-T (CAR-T) cell immunotherapy has produced dramatic responses in hematologic malignancies. One of the challenges in the field is the lack of a simple assay for the detection of CARs on the surface of immune effector cells. In this study, we describe a novel luciferase-based assay, termed Topanga Assay, for the detection of CAR expression. The assay utilizes a recombinant fusion protein, called Topanga reagent, generated by joining the extra-cellular domain of a CAR-target in frame with one of the marine luciferases or their engineered derivatives. The assay involves incubation of CAR expressing cells with the Topanga reagent, a few washes and measurement of luminescence. The assay can detect CARs comprising either immunoglobulin- or non-immunoglobulin-based antigen binding domains. We further demonstrate that addition of epitope tags to the Topanga reagent not only allows its convenient one step purification but also extends its use for detection of CAR cells using flow cytometry. However, crude supernatant containing the secreted Topanga reagent can be directly used in both luminescence and flow-cytometry based assays without prior protein purification. Our results demonstrate that the Topanga assay is a highly sensitive, specific, convenient, economical and versatile assay for the detection of CARs. |
| References: | Am J Clin Pathol. 2004 Feb;121(2):254-63. (PMID: 14983940) Mol Ther. 2005 Mar;11(3):435-43. (PMID: 15727940) Science. 2006 Oct 6;314(5796):126-9. (PMID: 16946036) Clin Cancer Res. 2008 May 1;14(9):2775-84. (PMID: 18451245) Blood. 2009 Aug 27;114(9):1974-86. (PMID: 19587380) J Thromb Haemost. 2010 Feb;8(2):301-10. (PMID: 19943873) Chem Biol. 2010 Jun 25;17(6):646-57. (PMID: 20609414) Blood. 2010 Nov 18;116(20):4099-102. (PMID: 20668228) J Biol Chem. 2011 Jun 17;286(24):21555-64. (PMID: 21531730) Mol Biol Evol. 2012 Jun;29(6):1669-81. (PMID: 22319154) J Transl Med. 2012 Feb 13;10:29. (PMID: 22330761) ACS Chem Biol. 2012 Nov 16;7(11):1848-57. (PMID: 22894855) J Transl Med. 2013 Jan 29;11:23. (PMID: 23360526) PLoS One. 2013;8(3):e57838. (PMID: 23469246) Clin Cancer Res. 2014 Aug 1;20(15):3989-4000. (PMID: 24677374) Haematologica. 2015 Mar;100(3):336-44. (PMID: 25480499) Biochem Biophys Res Commun. 2015 Jan 30;457(1):77-82. (PMID: 25543059) Leukemia. 2015 Aug;29(8):1637-47. (PMID: 25721896) Science. 2015 Oct 16;350(6258):aab4077. (PMID: 26405231) Ann Hematol. 2017 Aug;96(8):1407-1410. (PMID: 28577043) Mol Ther. 2017 Nov 1;25(11):2466-2476. (PMID: 28784559) Hum Gene Ther. 2017 Sep;28(9):726-736. (PMID: 28796529) Sci Rep. 2018 Jan 9;8(1):199. (PMID: 29317736) |
| معلومات مُعتمدة: | R01 DE025804 United States DE NIDCR NIH HHS |
| المشرفين على المادة: | 0 (Receptors, Antigen) 0 (Receptors, Antigen, T-Cell) 0 (Receptors, Chimeric Antigen) 0 (Recombinant Fusion Proteins) EC 1.13.12.- (Luciferases) |
| تواريخ الأحداث: | Date Created: 20190215 Date Completed: 20200825 Latest Revision: 20200825 |
| رمز التحديث: | 20221213 |
| مُعرف محوري في PubMed: | PMC6374361 |
| DOI: | 10.1038/s41598-018-38258-z |
| PMID: | 30760795 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2045-2322 |
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| DOI: | 10.1038/s41598-018-38258-z |