Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology
Background Protein microarrays have enormous potential as in vitro diagnostic tools stemming from the ability to miniaturize whilst generating maximum evaluation of diagnostically relevant information from minute amounts of sample. In this report, we present a method known as repeatable arrays of p...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BioMed Central
2015
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| Online Access: | http://eprints.intimal.edu.my/16/ http://eprints.intimal.edu.my/16/1/Development%20of%20Repeatable%20Arrays%20of%20Proteins%20using%20Immobilized%20DNA%20Microplate%20%28RAPID-M%29%20Technology.pdf |
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| author | Ashaari, Nur Suhanawati Ramarad, Suganti Khairuddin, Dzulaikha Mat Akhir, Nor Azurah Yuka, Hara Mahadi, Nor Muhammad Rahmah, Mohamed Nathan, Sheila |
| author_facet | Ashaari, Nur Suhanawati Ramarad, Suganti Khairuddin, Dzulaikha Mat Akhir, Nor Azurah Yuka, Hara Mahadi, Nor Muhammad Rahmah, Mohamed Nathan, Sheila |
| author_sort | Ashaari, Nur Suhanawati |
| building | INTI Institutional Repository |
| collection | Online Access |
| description | Background
Protein microarrays have enormous potential as in vitro diagnostic tools stemming from the ability to miniaturize whilst generating maximum evaluation of diagnostically relevant information from minute amounts of sample. In this report, we present a method known as repeatable arrays of proteins using immobilized DNA microplates (RAPID-M) for high-throughput in situ protein microarray fabrication. The RAPID-M technology comprises of cell-free expression using immobilized DNA templates and in situ protein purification onto standard microarray slides.
Results
To demonstrate proof-of-concept, the repeatable protein arrays developed using our RAPID-M technology utilized green fluorescent protein (GFP) and a bacterial outer membrane protein (OmpA) as the proteins of interest for microarray fabrication. Cell-free expression of OmpA and GFP proteins using beads-immobilized DNA yielded protein bands with the expected molecular sizes of 27 and 30 kDa, respectively. We demonstrate that the beads-immobilized DNA remained stable for at least four cycles of cell-free expression. The OmpA and GFP proteins were still functional after in situ purification on the Ni–NTA microarray slide.
Conclusion
The RAPID-M platform for protein microarray fabrication of two different representative proteins was successfully developed. |
| first_indexed | 2025-11-14T11:50:10Z |
| format | Article |
| id | intimal-16 |
| institution | INTI International University |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T11:50:10Z |
| publishDate | 2015 |
| publisher | BioMed Central |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | intimal-162017-03-03T01:49:03Z http://eprints.intimal.edu.my/16/ Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology Ashaari, Nur Suhanawati Ramarad, Suganti Khairuddin, Dzulaikha Mat Akhir, Nor Azurah Yuka, Hara Mahadi, Nor Muhammad Rahmah, Mohamed Nathan, Sheila TP Chemical technology Background Protein microarrays have enormous potential as in vitro diagnostic tools stemming from the ability to miniaturize whilst generating maximum evaluation of diagnostically relevant information from minute amounts of sample. In this report, we present a method known as repeatable arrays of proteins using immobilized DNA microplates (RAPID-M) for high-throughput in situ protein microarray fabrication. The RAPID-M technology comprises of cell-free expression using immobilized DNA templates and in situ protein purification onto standard microarray slides. Results To demonstrate proof-of-concept, the repeatable protein arrays developed using our RAPID-M technology utilized green fluorescent protein (GFP) and a bacterial outer membrane protein (OmpA) as the proteins of interest for microarray fabrication. Cell-free expression of OmpA and GFP proteins using beads-immobilized DNA yielded protein bands with the expected molecular sizes of 27 and 30 kDa, respectively. We demonstrate that the beads-immobilized DNA remained stable for at least four cycles of cell-free expression. The OmpA and GFP proteins were still functional after in situ purification on the Ni–NTA microarray slide. Conclusion The RAPID-M platform for protein microarray fabrication of two different representative proteins was successfully developed. BioMed Central 2015 Article PeerReviewed text en http://eprints.intimal.edu.my/16/1/Development%20of%20Repeatable%20Arrays%20of%20Proteins%20using%20Immobilized%20DNA%20Microplate%20%28RAPID-M%29%20Technology.pdf Ashaari, Nur Suhanawati and Ramarad, Suganti and Khairuddin, Dzulaikha and Mat Akhir, Nor Azurah and Yuka, Hara and Mahadi, Nor Muhammad and Rahmah, Mohamed and Nathan, Sheila (2015) Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology. BMC Research Notes, 8 (1). ISSN 1756-0500 http://www.biomedcentral.com/ 10.1186/s13104-015-1637-3 |
| spellingShingle | TP Chemical technology Ashaari, Nur Suhanawati Ramarad, Suganti Khairuddin, Dzulaikha Mat Akhir, Nor Azurah Yuka, Hara Mahadi, Nor Muhammad Rahmah, Mohamed Nathan, Sheila Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology |
| title | Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology |
| title_full | Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology |
| title_fullStr | Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology |
| title_full_unstemmed | Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology |
| title_short | Development of Repeatable Arrays of Proteins using Immobilized DNA Microplate (RAPID-M) Technology |
| title_sort | development of repeatable arrays of proteins using immobilized dna microplate (rapid-m) technology |
| topic | TP Chemical technology |
| url | http://eprints.intimal.edu.my/16/ http://eprints.intimal.edu.my/16/ http://eprints.intimal.edu.my/16/ http://eprints.intimal.edu.my/16/1/Development%20of%20Repeatable%20Arrays%20of%20Proteins%20using%20Immobilized%20DNA%20Microplate%20%28RAPID-M%29%20Technology.pdf |