دورية أكاديمية
Multiplexed DNA-Directed Patterning of Antibodies for Applications in Cell Subpopulation Analysis.
| العنوان: | Multiplexed DNA-Directed Patterning of Antibodies for Applications in Cell Subpopulation Analysis. |
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| المؤلفون: | Kozminsky M; California Institute of Quantitative Biosciences, University of California, Berkeley, 174 Stanley Hall, Berkeley, California 94720, United States., Scheideler OJ; The UC Berkeley-UC San Francisco Graduate Program in Bioengineering, University of California, Berkeley, 306 Stanley Hall, Berkeley, California 94720, United States., Li B; The UC Berkeley-UC San Francisco Graduate Program in Bioengineering, University of California, Berkeley, 306 Stanley Hall, Berkeley, California 94720, United States., Liu NK; Department of Mechanical Engineering, University of California, Berkeley, 5118 Etcheverry Hall, Berkeley, California 94720, United States., Sohn LL; California Institute of Quantitative Biosciences, University of California, Berkeley, 174 Stanley Hall, Berkeley, California 94720, United States.; The UC Berkeley-UC San Francisco Graduate Program in Bioengineering, University of California, Berkeley, 306 Stanley Hall, Berkeley, California 94720, United States.; Department of Mechanical Engineering, University of California, Berkeley, 5118 Etcheverry Hall, Berkeley, California 94720, United States. |
| المصدر: | ACS applied materials & interfaces [ACS Appl Mater Interfaces] 2021 Oct 06; Vol. 13 (39), pp. 46421-46430. Date of Electronic Publication: 2021 Sep 21. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 101504991 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1944-8252 (Electronic) Linking ISSN: 19448244 NLM ISO Abbreviation: ACS Appl Mater Interfaces Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, D.C. : American Chemical Society |
| مواضيع طبية MeSH: | Antibodies/*immunology , Cell Separation/*methods , DNA/*chemistry, Antigens, CD/immunology ; Antigens, CD/metabolism ; Biomarkers/metabolism ; Cadherins/immunology ; Cadherins/metabolism ; Cell Line, Tumor ; Cell Separation/instrumentation ; Epithelial-Mesenchymal Transition/physiology ; Humans ; Immunoassay/instrumentation ; Immunoassay/methods ; Integrin beta Chains/immunology ; Integrin beta Chains/metabolism ; Lab-On-A-Chip Devices ; Microfluidic Analytical Techniques/instrumentation ; Microfluidic Analytical Techniques/methods ; Oligodeoxyribonucleotides/chemistry ; Proof of Concept Study |
| مستخلص: | Antibodies provide the functional biospecificity that has enabled the development of sensors, diagnostic tools, and assays in both laboratory and clinical settings. However, as multimarker screening becomes increasingly necessary due to the heterogeneity and complexity of human pathology, new methods must be developed that are capable of coordinating the precise assembly of multiple, distinct antibodies. To address this technological challenge, we engineered a bottom-up, high-throughput method in which DNA patterns, comprising unique 20-base pair oligonucleotides, are patterned onto a substrate using photolithography. These microfabricated surface patterns are programmed to hybridize with, and instruct the multiplexed assembly of, antibodies conjugated with the complementary DNA strands. We demonstrate that this simple, yet robust, approach preserves the antibody-binding functionality in two common applications: antibody-based cell capture and label-free surface marker screening. Using a simple proof-of-concept capture device, we achieved high purity separation of a breast cancer cell line, MCF-7, from a blood cell line, Jurkat, with capture purities of 77.4% and 96.6% when using antibodies specific for the respective cell types. We also show that antigen-antibody interactions slow cell trajectories in flow in the next-generation microfluidic node-pore sensing (NPS) device, enabling the differentiation of MCF-7 and Jurkat cells based on EpCAM surface-marker expression. Finally, we use a next-generation NPS device patterned with antibodies against E-cadherin, N-cadherin, and β-integrin-three markers that are associated with epithelial-mesenchymal transitions-to perform label-free surface marker screening of MCF10A, MCF-7, and Hs 578T breast epithelial cells. Our high-throughput, highly versatile technique enables rapid development of customized, antibody-based assays across a host of diverse diseases and research thrusts. |
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| معلومات مُعتمدة: | F32 CA243354 United States CA NCI NIH HHS; R01 CA190843 United States CA NCI NIH HHS; R01 EB024989 United States EB NIBIB NIH HHS |
| فهرسة مساهمة: | Keywords: DNA-directed patterning; antibody patterning; breast cancer; cell capture; cell surface markers; node-pore sensing; photolithography |
| المشرفين على المادة: | 0 (Antibodies) 0 (Antigens, CD) 0 (Biomarkers) 0 (CDH1 protein, human) 0 (CDH2 protein, human) 0 (Cadherins) 0 (Integrin beta Chains) 0 (Oligodeoxyribonucleotides) 9007-49-2 (DNA) |
| تواريخ الأحداث: | Date Created: 20210921 Date Completed: 20220131 Latest Revision: 20220216 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC8817232 |
| DOI: | 10.1021/acsami.1c15047 |
| PMID: | 34546726 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1944-8252 |
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| DOI: | 10.1021/acsami.1c15047 |