Plasma Activation of Microplates Optimized for One-Step Reagent-Free Immobilization of DNA and Protein
Field | Value | Language |
dc.contributor.author | Coffi Dit Gleize, Kanako | |
dc.contributor.author | Tran, Clara T. H. | |
dc.contributor.author | Waterhouse, Anna | |
dc.contributor.author | Bilek, Marcela M. M. | |
dc.contributor.author | Wickham, Shelley F. J. | |
dc.date.accessioned | 2024-03-15T05:38:08Z | |
dc.date.available | 2024-03-15T05:38:08Z | |
dc.date.issued | 2022 | en_AU |
dc.identifier.uri | https://hdl.handle.net/2123/32377 | |
dc.description.abstract | Activated microplates are widely used in biological assays and cell culture to immobilize biomolecules, either through passive physical adsorption or covalent cross-linking. Covalent attachment gives greater stability in complex biological mixtures. However, current multistep chemical activation methods add complexity and cost, require specific functional groups, and can introduce cytotoxic chemicals that affect downstream cellular applications. Here, we show a method for one-step linker-free activation of microplates by energetic ions from plasma for covalent immobilization of DNA and protein. Two types of energetic ion plasma treatment were shown to be effective: plasma immersion ion implantation (PIII) and plasma-activated coating (PAC). This is the first time that PIII and PAC have been reported in microwell plates with nonflat geometry. We confirm that the plasma treatment generates radical-activated surfaces at the bottom of wells despite potential shadowing from the walls. Comprehensive surface characterization studies were used to compare the PIII and PAC microplate surface composition, wettability, radical density, optical properties, stability, and biomolecule immobilization density. PAC plates were found to have more nitrogen and lower radical density and were more hydrophobic and more stable over 3 months than PIII plates. Optimal conditions were obtained for high-density DNA (PAC, 0 or 21% nitrogen, pH 3–4) and streptavidin (PAC, 21% nitrogen, pH 5–7) binding while retaining optical properties required for typical high-throughput biochemical microplate assays, such as low autofluorescence and high transparency. DNA hybridization and protein activity of immobilized molecules were confirmed. We show that PAC activation allows for high-density covalent immobilization of functional DNA and protein in a single step on both 96- and 384-well plates without specific linker chemistry. These microplates could be used in the future to bind other user-selected ligands in a wide range of applications, for example, for solid phase polymerase chain reaction and stem cell culture and differentiation. | en_AU |
dc.language.iso | en | en_AU |
dc.publisher | ACS Publications | en_AU |
dc.relation.ispartof | Langmuir | en_AU |
dc.rights | Copyright All Rights Reserved | en_AU |
dc.title | Plasma Activation of Microplates Optimized for One-Step Reagent-Free Immobilization of DNA and Protein | en_AU |
dc.type | Article | en_AU |
dc.identifier.doi | 10.1021/acs.langmuir.2c02573 | |
dc.type.pubtype | Author accepted manuscript | en_AU |
dc.relation.arc | DE180101635 | |
dc.relation.arc | FL190100216 | |
dc.relation.other | Westpac Research Fellowship | |
dc.relation.other | The University of Sydney | |
dc.relation.other | The Heart Research Institute | |
dc.relation.other | The University of Sydney Nano Institute | |
usyd.faculty | SeS faculties schools::Faculty of Science::School of Chemistry | en_AU |
usyd.faculty | SeS faculties schools::Faculty of Science::School of Physics | en_AU |
usyd.faculty | SeS faculties schools::Faculty of Medicine and Health::School of Medical Sciences | en_AU |
usyd.faculty | SeS faculties schools::The University of Sydney Multidisciplinary Centres and Institutes ::The University of Sydney Nano Institute | en_AU |
usyd.faculty | SeS faculties schools::The University of Sydney Multidisciplinary Centres and Institutes ::Charles Perkins Centre | en_AU |
usyd.faculty | The Heart Research Institute, The University of Sydney, Newtown 2042, Australia | en_AU |
usyd.faculty | SeS faculties schools::Faculty of Engineering::School of Biomedical Engineering | en_AU |
usyd.citation.volume | 39 | en_AU |
usyd.citation.issue | 1 | en_AU |
usyd.citation.spage | 343 | en_AU |
usyd.citation.epage | 356 | en_AU |
workflow.metadata.only | No | en_AU |
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